EP4240408A1 - Verfahren zur behandlung eines tumors mit einer kombination aus einem il-7-protein und einem nukleotidimpfstoff - Google Patents

Verfahren zur behandlung eines tumors mit einer kombination aus einem il-7-protein und einem nukleotidimpfstoff

Info

Publication number
EP4240408A1
EP4240408A1 EP21830359.2A EP21830359A EP4240408A1 EP 4240408 A1 EP4240408 A1 EP 4240408A1 EP 21830359 A EP21830359 A EP 21830359A EP 4240408 A1 EP4240408 A1 EP 4240408A1
Authority
EP
European Patent Office
Prior art keywords
days
glycine
administered
protein
methionine
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP21830359.2A
Other languages
English (en)
French (fr)
Inventor
Byung Ha Lee
Donghoon Choi
William GILLANDERS
Ina CHEN
Simon Peter GOEDEGEBUURE
Lijin LI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Washington University in St Louis WUSTL
Neoimmunetech Inc
Original Assignee
Washington University in St Louis WUSTL
Neoimmunetech Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Washington University in St Louis WUSTL, Neoimmunetech Inc filed Critical Washington University in St Louis WUSTL
Publication of EP4240408A1 publication Critical patent/EP4240408A1/de
Pending legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/20—Interleukins [IL]
    • A61K38/2046—IL-7
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001136—Cytokines
    • A61K39/00114—Interleukins [IL]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
    • A61K47/6813—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin the drug being a peptidic cytokine, e.g. an interleukin or interferon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55522—Cytokines; Lymphokines; Interferons
    • A61K2039/55527—Interleukins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00

Definitions

  • a method of treating a tumor in a subject in need thereof comprising administering to the subject a nucleotide vaccine encoding a tumor antigen in combination with an interleukin-7 (IL-7), wherein the administration of the nucleotide vaccine induces a tumor-specific T cell immune response, and wherein the IL-7 is administered to the subject within about 14 days, within about 13 days, within about 12 days, within about 11 days, within about 10 days, within about nine days, within about eight days, within about seven days, within about six days, within about five days, within about four days, within about three days, within about two days, or within about one day of the nucleotide vaccine administration.
  • the IL-7 is administered within about seven days of the nucleotide vaccine administration.
  • the IL-7 and the nucleotide vaccine are administered to the subject concurrently.
  • a method of treating a tumor in a subject in need thereof comprising administering to the subject a nucleotide vaccine encoding a tumor antigen in combination with an interleukin-7 (IL-7), wherein the administration of the nucleotide vaccine induces a tumor-specific T cell immune response, and wherein the IL-7 is administered to the subject after a peak expansion phase of the tumor-specific T cell immune response.
  • IL-7 interleukin-7
  • a tumor volume is reduced in the subject after the administration compared to a reference (e.g., corresponding value in a subject that received either IL-7 alone or nucleotide vaccine alone).
  • the tumor volume is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% after the administration compared to the reference.
  • nucleotide vaccine encoding a tumor antigen in combination with an interleukin-7 (IL-7), wherein the administration of the nucleotide vaccine induces a tumor-specific T cell immune response, and wherein the nucleotide vaccine, the IL-7, or both the nucleotide vaccine and the IL-7 are administered to the subject prior to the occurrence of the tumor.
  • IL-7 interleukin-7
  • the IL-7 is administered to the subject within about 14 days, within about 13 days, within about 12 days, within about 11 days, within about 10 days, within about nine days, within about eight days, within about seven days, within about six days, within about five days, within about four days, within about three days, within about two days, or within about one day of the nucleotide vaccine administration. In some aspects, the IL-7 is administered to the subject within about seven days of the nucleotide vaccine administration. In some aspects, the IL-7 and the nucleotide vaccine are administered to the subject concurrently.
  • Also provided herein is a method of prolonging a tumor-specific T cell immune response in a subject in need thereof, comprising administering to the subject a nucleotide vaccine encoding a tumor antigen in combination with an interleukin-7 (IL-7), wherein the administration of the nucleotide vaccine induces a tumor-specific T cell immune response, and wherein the IL-7 is administered to the subject after a peak expansion phase of the tumorspecific T cell immune response.
  • IL-7 interleukin-7
  • Present disclosure further provides a method of prolonging a tumor-specific T cell immune response in a subject in need thereof, comprising administering to the subject a nucleotide vaccine encoding a tumor antigen in combination with an interleukin-7 (IL-7), wherein the administration of the nucleotide vaccine induces a tumor-specific T cell immune response, and wherein the IL-7 is administered to the subject within about 14 days, within about 13 days, within about 12 days, within about 11 days, within about 10 days, within about nine days, within about eight days, within about seven days, within about six days, within about five days, within about four days, within about three days, within about two days, or within about one day of the nucleotide vaccine administration.
  • the IL-7 is administered within about seven days of the nucleotide vaccine administration.
  • the IL-7 and the nucleotide vaccine are administered to the subject concurrently.
  • the administration of IL-7 increases a survival of tumor-specific T cells during a contraction phase of the tumor-specific T cell immune response, compared to a reference (e.g., corresponding value in a subject that received either IL-7 alone or nucleotide vaccine alone).
  • the survival of tumor-specific T cells during the contraction phase is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8- fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the reference.
  • the administration of IL-7 increases a number of tumor-specific T cells during a contraction phase of the tumor-specific T cell immune response, compared to a reference (e.g., corresponding value in a subject that received either IL-7 alone or nucleotide vaccine alone).
  • the number of tumor-specific T cells during the contraction phase of the tumor-specific T cell immune response is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the reference.
  • TCR T-cell receptor
  • IL-7 interleukin-7
  • a method of expanding a T-cell receptor (TCR) repertoire of a tumor-specific T cell immune response in a subject in need thereof comprising administering to the subject a nucleotide vaccine encoding a tumor antigen in combination with an interleukin-7 (IL-7), wherein the administration of the nucleotide vaccine induces a tumorspecific T cell immune response against one or more epitopes of the tumor antigen, and wherein the IL-7 is administered to the subject within about 14 days, within about 13 days, within about 12 days, within about 11 days, within about 10 days, within about nine days, within about eight days, within about seven days, within about six days, within about five days, within about four days, within about three days, within about two days, or within about one day of the nucleotide vaccine administration.
  • the IL-7 is administered within about seven days of the nucleotide vaccine administration.
  • the IL-7 and the nucleotide vaccine are administered to the subject concurrently.
  • the administration increases the number of epitopes against which the tumor-specific T cell immune response is induced, compared to a reference (e.g., corresponding value in a subject that received either IL-7 alone or nucleotide vaccine alone).
  • the number of epitopes against which the tumor-specific T cell immune response is induced is increased by at least about 1-fold, at least about 2-fold, at least about 3- fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the reference.
  • the tumor antigen is derived from a breast cancer and the epitopes are selected from Lrrc27, Plekhol, Pttgl, Xpo4, Exoc4, Pank3, TmemlOl, Map3k6, Met, BC057079, Histlh3e, Prkagl, Neil3, or combinations thereof.
  • the administration induces a tumor-specific T cell immune response to at least about three, at least about four, at least about five, at least about six, at least about seven, at least about eight, at least about nine, at least about 10, at least about 11, at least about 12, or at least about 13 or more epitopes on the tumor antigen.
  • Present disclosure further provides a method of increasing a T cell immune response against a subdominant epitope of a tumor antigen in a subject in need thereof, comprising administering to the subject a nucleotide vaccine encoding a tumor antigen, which comprises the subdominant epitope, in combination with an interleukin-7 (IL-7), wherein the IL-7 is administered to the subject after a peak expansion phase of the tumor-specific T cell immune response.
  • IL-7 interleukin-7
  • a method of increasing a T cell immune response against a subdominant epitope of a tumor antigen in a subject in need thereof comprising administering to the subject a nucleotide vaccine encoding a tumor antigen, which comprises the subdominant epitope, in combination with an interleukin-7 (IL-7), wherein the IL-7 is administered to the subject within about 14 days, within about 13 days, within about 12 days, within about 11 days, within about 10 days, within about nine days, within about eight days, within about seven days, within about six days, within about five days, within about four days, within about three days, within about two days, or within about one day of the nucleotide vaccine administration.
  • the IL-7 is administered within about seven days of the nucleotide vaccine administration.
  • the IL-7 and the nucleotide vaccine are administered to the subject concurrently.
  • a T cell immune response against a subdominant epitope of a tumor antigen is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8- fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to a reference (e.g., corresponding value in a subject that received an IL-7 alone or nucleotide vaccine alone).
  • a reference e.g., corresponding value in a subject that received an IL-7 alone or nucleotide vaccine alone.
  • the peak expansion phase of the tumor-specific T cell immune response occurs at about seven days, about eight days, about nine days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, or about 15 days after an initial administration of the nucleotide vaccine. In some aspects, the peak expansion phase of the tumor-specific T cell immune response occurs at about 11 days after an initial administration of the nucleotide vaccine.
  • the IL-7 is administered at least about one day, about two days, about three days, about four days, about five days, about six days, about seven days, about eight days, about nine days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, or about 30 days or more after the peak expansion phase of the tumor-specific T cell immune response.
  • the IL-7 is administered at about two days after the peak expansion phase of the tumorspecific T cell immune response.
  • the IL-7 is administered at a dose between about 5 mg/kg and about 15 mg/kg. In some aspects, the IL-7 is administered at a dose of about 5 mg/kg. In some aspects, the IL-7 is administered at a dose of between about 20 pg/kg and about 600 pg/kg. In certain aspects, the IL-7 protein is administered at a dose of about 20 pg/kg, about 60 pg/kg, about 120 pg/kg, about 240 pg/kg, about 480 pg/kg, or about 600 pg/kg.
  • the IL-7 protein is administered at a dose greater than about 600 pg/kg, greater than about 700 pg/kg, greater than about 800 pg/kg, greater than about 900 pg/kg, greater than about 1,000 pg/kg, greater than about 1,100 pg/kg, greater than about 1,200 pg/kg, greater than about 1,300 pg/kg, greater than about 1,400 pg/kg, greater than about 1,500 pg/kg, greater than about 1,600 pg/kg, greater than about 1,700 pg/kg, greater than about 1,800 pg/kg, greater than about 1,900 pg/kg, or greater than about 2,000 pg/kg.
  • the IL-7 protein is administered at a dose of between about 610 pg/kg and about 1,200 pg/kg, between about 650 pg/kg and about 1,200 pg/kg, between about 700 pg/kg and about 1,200 pg/kg, between about 750 pg/kg and about 1,200 pg/kg, between about 800 pg/kg and about 1,200 pg/kg, between about 850 pg/kg and about 1,200 pg/kg, between about 900 pg/kg and about 1,200 pg/kg, between about 950 pg/kg and about 1,200 pg/kg, between about 1,000 pg/kg and about 1,200 pg/kg, between about 1,050 pg/kg and about 1,200 pg/kg, between about 1,100 pg/kg and about 1,200 pg/kg, between about 1,200 pg/kg and about 2,000 pg/kg, between about 1,300 pg/kg
  • the IL-7 protein is administered at a dose of between about 700 pg/kg and about 900 pg/kg, between about 750 pg/kg and about 950 pg/kg, between about 700 pg/kg and about 850 pg/kg, between about 750 pg/kg and about 850 pg/kg, between about 700 pg/kg and about 800 pg/kg, between about 800 pg/kg and about 900 pg/kg, between about 750 pg/kg and about 850 pg/kg, or between about 850 pg/kg and about 950 pg/kg.
  • the IL-7 protein is administered at a dose of about 650 pg/kg, about 680 pg/kg, about 700 pg/kg, about 720 pg/kg, about 740 pg/kg, about 750 pg/kg, about 760 pg/kg, about 780 pg/kg, about 800 pg/kg, about 820 pg/kg, about 840 pg/kg, about 850 pg/kg, about 860 pg/kg, about 880 pg/kg, about 900 pg/kg, about 920 pg/kg, about 940 pg/kg, about 950 pg/kg, about 960 pg/kg, about 980 pg/kg, about 1,000 pg/kg, about 1,100 pg/kg, about 1200 pg/kg, about 1,300 pg/kg, about 1,400 pg/kg, about 1,440 pg/kg, about 1,500
  • the IL-7 is administered at a dosing frequency of about once a week, about once in two weeks, about once in three weeks, about once in four weeks, about once in five weeks, about once in six weeks, about once in seven weeks, about once in eight weeks, about once in nine weeks, about once in 10 weeks, about once in 11 weeks, or about once in 12 weeks.
  • the nucleotide vaccine comprises a DNA vaccine, mRNA vaccine, or both.
  • the nucleotide vaccine is a DNA vaccine.
  • the IL-7 is administered as a protein (IL-7 protein), nucleic acid encoding the IL-7 protein, or both.
  • the subject is a human.
  • the IL-7 protein of a method disclosed herein is not a wild-type IL-7.
  • the IL-7 protein is a fusion protein.
  • the IL-7 protein comprises an oligopeptide consisting of 1 to 10 amino acid residues.
  • oligopeptide comprises methionine (M), glycine (G), methionine-methionine (MM), glycineglycine (GG), methionine-glycine (MG), glycine-methionine (GM), methionine-methionine- methionine (MMM), methionine-methionine-glycine (MMG), methionine-glycine-methionine (MGM), methionine-glycine-methionine (GMM), methionine-glycine-glycine (MGG), glycine-methionine-glycine (GMG), glycine-glycine-methionine (GGM), glycine-glycine-methionine (GGM), glycine-glycine-methionine (GGM), glycine-glycine-methionine (GGM), glycine-glycine-methionine (GGM), g
  • the IL-7 protein comprises a half-life extending moiety.
  • the half-life extending moiety comprises an Fc, albumin, an albumin-binding polypeptide, Pro/Ala/Ser (PAS), a C-terminal peptide (CTP) of the P subunit of human chorionic gonadotropin, polyethylene glycol (PEG), long unstructured hydrophilic sequences of amino acids (XTEN), hydroxyethyl starch (HES), an albumin-binding small molecule, or a combination thereof.
  • the half-life extending moiety is an Fc.
  • the Fc is a hybrid Fc, comprising a hinge region, a CH2 domain, and a CH3 domain, wherein the hinge region comprises a human IgD hinge region, wherein the CH2 domain comprises a part of human IgD CH2 domain and a part of human IgG4 CH2 domain, and wherein the CH3 domain comprises a part of human IgG4 CH3 domain.
  • the IL-7 protein comprises an amino acid sequence having a sequence identity of at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% to SEQ ID NOs: 1-6 and 15-25.
  • the IL-7 is administered to the subject parenthetically, intramuscularly, subcutaneously, ophthalmic, intravenously, intraperitoneally, intradermally, intraorbitally, intracerebrally, intracranially, intraspinally, intraventricular, intrathecally, intraci stemally, intracapsularly, intratumorally, or any combination thereof.
  • the nucleotide vaccine is administered to the subject parenthetically, intramuscularly, cutaneously, subcutaneously, ophthalmic, intravenously, intraperitoneally, intradermally, intraorbitally, intracerebrally, intracranially, intraspinally, intraventricular, intrathecally, intraci stemally, intracapsularly, intratumorally, or any combination thereof.
  • methods provided herein further comprises administering at least one additional therapeutic agent to the subject.
  • the tumor antigen comprises guanylate cyclase C (GC-C), epidermal growth factor receptor (EGFR or erbB-1), human epidermal growth factor receptor 2 (HER2 or erbB2), erbB-3, erbB-4, MUC-1, melanoma- associated chondroitin sulfate proteoglycan (MCSP), mesothelin (MSLN), folate receptor 1 (FOLR1), CD4, CD 19, CD20, CD22, CD30, CD33, CD38, CD44, CD44v6, CD44v7/8, CD70, CD123, CD138, CD171, CEA, CSPG4, CXCR5, c-Met, HERV-envelope protein, eriostin, Bigh3, SPARC, BCR, CD79, CD37, EGFRvIII, EGP2, EGP40, IGFr, L1CAM, AXL, Tissue Factor (TF), CD
  • GC-C guanylate cyclase C
  • the tumor antigen is derived from a cancer comprising a breast cancer, head and neck cancer, uterine cancer, brain cancer, skin cancer, renal cancer, lung cancer, colorectal cancer, prostate cancer, liver cancer, bladder cancer, kidney cancer, pancreatic cancer, thyroid cancer, esophageal cancer, eye cancer, stomach (gastric) cancer, gastrointestinal cancer, ovarian cancer, carcinoma, sarcoma, leukemia, lymphoma, myeloma, or a combination thereof.
  • a cancer comprising a breast cancer, head and neck cancer, uterine cancer, brain cancer, skin cancer, renal cancer, lung cancer, colorectal cancer, prostate cancer, liver cancer, bladder cancer, kidney cancer, pancreatic cancer, thyroid cancer, esophageal cancer, eye cancer, stomach (gastric) cancer, gastrointestinal cancer, ovarian cancer, carcinoma, sarcoma, leukemia, lymphoma, myeloma, or a combination thereof.
  • FIGs. 1A, IB, 1C, ID, IE, and IF show the effect of IL-7 administration at different time points after DNA vaccine administration.
  • FIG. 1A provides a schematic of the experimental design. As shown, animals received three immunizations of the DNA vaccine at a dosing frequency of once every three days. The IL-7 was administered to the animals at either day 4 or day 13 post initial DNA vaccine administration. The peak tumor-specific T cell immune response was observed at day 11 post initial DNA vaccine administration.
  • FIG. IB provides images comparing the spleen size at day 11 in animals that received (i) the control vector or DNA vaccine alone (left picture) or (ii) DNA vaccine + IL-7 at day 4 post initial DNA administration (right picture).
  • 1C and ID provide comparison of the frequency (# of IFN-y-producing T cells / 10 6 total splenocytes) and total number per spleen of tumor-specific T cells (i.e., specific to one of the following epitopes: Lrrc27, Plekhol, or Pttgl) at day 11 post initial DNA vaccine administration from the different treatment groups.
  • the groups shown are as follows (from left to right) (i) control vector alone (Gl); (ii) DNA vaccine alone (G2); and (iii) DNA vaccine + IL-7 at day 4 post initial DNA administration (G3).
  • IE and IF provide comparison of the frequency (# of IFN-y- producing T cells / 10 6 total splenocytes) and total number of tumor-specific T cells (i.e., specific to one of the following epitopes: Lrrc27, Plekhol, or Pttgl) at day 20 post initial DNA vaccine administration from the following treatment groups: (i) control vector alone (Gl); (ii) DNA vaccine alone (G2); (iii) DNA vaccine + IL-7 at day 4 post initial DNA administration (G3); and (iv) DNA vaccine + IL-7 at day 13 post initial DNA administration (G4).
  • the groups shown are as follows (from left to right) (i) control vector alone (Gl); (ii) DNA vaccine alone (G2); (iii) DNA vaccine + IL-7 at day 4 post initial DNA administration (G3); and (iv) DNA vaccine + IL-7 at day 13 post initial DNA administration (G4).
  • FIGs. 2A, 2B, and 2C show the effect of IL-7 dosage on tumor-specific T cell immune response after DNA vaccine administration.
  • FIG. 2A provides a schematic of the experimental design. As shown, animals received three immunizations of the DNA vaccine at a dosing frequency of once every three days. The IL-7 was administered to the animals at day 13 post initial DNA vaccine administration at one of the following doses: 5, 10, or 15 mg/kg. FIGs.
  • 2B and 2C provide comparison of the frequency (# of IFN-y-producing T cells / 10 6 total splenocytes) of tumor-specific T cells (i.e., specific to one of the following epitopes: Lrrc27, Plekhol, or Pttgl) at day 20 post initial DNA administration in the spleen and lymph nodes, respectively.
  • tumor-specific T cells i.e., specific to one of the following epitopes: Lrrc27, Plekhol, or Pttgl
  • the groups shown are as follows (from left to right) (i) control vector alone (Gl); (ii) DNA vaccine alone (G2); (iii) DNA vaccine + 5 mg/kg of IL-7 (G3); (iv) DNA vaccine + 10 mg/kg of IL-7 (G4); and (v) DNA vaccine + 15 mg/kg of IL-7 (G5).
  • FIGs. 3A and 3B show the anti-tumor effects of a nucleotide vaccine and IL-7 combination therapy disclosed herein.
  • FIG. 3A provides a schematic of the experimental design. As shown, animals received three immunizations of the DNA vaccine at a dosing frequency of once every three days. At day 8 post initial DNA immunization, animals were implanted subcutaneously with E0771 tumor cells (5 x 10 5 cells/mouse). Peak tumor-specific T cell immune response was observed at about day 10 post initial DNA immunization. The IL- 7 was administered to the animals at day 13 post initial DNA vaccine administration.
  • FIG. 3B provides a comparison of the tumor volume in animals from the different treatment groups at various time points post initial DNA vaccine administration. The groups shown include: (i) control vector only (circle); (ii) DNA vaccine only (square); and (iii) DNA vaccine + IL-7 (triangle).
  • FIGs. 4A and 4B show the effect of a nucleotide vaccine and IL-7 combination therapy described herein on T cell-mediated cytotoxicity as measured using an in vivo CTL assay.
  • FIG. 4A provides a schematic of the experimental design.
  • FIG. 4B provides a comparison of the percent killing of the neoantigen-pulsed splenocytes in animals from the different treatment groups at days 22, 34, 41, and 51 post initial immunization.
  • the different treatment groups included: (i) control vector only ("vector”); (ii) DNA vaccine only ("nAg”); (iii) IL-7 protein alone (“IL-7”); and (iv) DNA vaccine + IL-7 protein ("nAg + IL-7"). Percent killing of the pulsed splenocytes is shown normalized to the un-pulsed splenocyte control.
  • FIGs. 5A and 5B show the anti-tumor effects of a nucleotide vaccine and IL-7 combination therapy when administered after the occurrence of a tumor.
  • FIG. 5A provides a schematic of the experimental design.
  • FIG. 5B provides comparison of the frequency (# of IFN-y-producing T cells / 10 6 total splenocytes) of tumor-specific T cells (i.e., specific to one of the following: Lrrc27, Plekhol, Pttgl, no peptide ("media”) - left to right in each of the treatment groups shown) at days 20 post animal randomization.
  • the different treatment groups are shown along the x-axis and further described in Example 6.
  • FIGs. 6A and 6B show the anti-tumor effects of a nucleotide vaccine and IL-7 combination therapy as a prophylactic vaccine.
  • FIG. 6A provides a schematic of the experimental design.
  • FIG. 6B provides a comparison of tumor volume in animals from the different treatment groups at various time points post tumor inoculation.
  • a or “an” entity refers to one or more of that entity; for example, “an antibody,” is understood to represent one or more antibodies.
  • an antibody is understood to represent one or more antibodies.
  • the terms “a” (or “an”), “one or more,” and “at least one” can be used interchangeably herein.
  • administering refers to the physical introduction of a therapeutic agent or a composition comprising a therapeutic agent to a subject, using any of the various methods and delivery systems known to those skilled in the art.
  • the different routes of administration for a therapeutic agent described herein include intravenous, intraperitoneal, intramuscular, cutaneous, subcutaneous, spinal, intratumorally, or other parenteral routes of administration, for example by injection or infusion.
  • parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, intratracheal, pulmonary, subcuticular, intraarticular, subcapsular, subarachnoid, intraventricle, intravitreal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation.
  • a therapeutic agent described herein can be administered via a non-parenteral route, such as a topical, epidermal, or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually, or topically.
  • Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.
  • the term "antigen” refers to any natural or synthetic immunogenic substance, such as a protein, peptide, or hapten.
  • the antigen comprises a tumor antigen.
  • tumor antigen refers to an antigen that is uniquely or differentially expressed on a tumor cell compared to normal healthy cells.
  • epitope refers to a set of amino acid residues that is involved in recognition by a particular immunoglobulin, or in the context of T cells, those residues necessary or recognition by T cell receptor proteins and/or major histocompatibility complex (MHC) receptors (e.g., site on a tumor antigen to which a tumor-specific T cell can recognize and target).
  • MHC major histocompatibility complex
  • an epitope is the collective features of a molecule, such as primary, secondary and tertiary peptide structure, and charge, that together form a site recognized by an immunoglobulin, T cell receptor, or HLA molecule.
  • Epitopes can be formed both from contiguous amino acids (linear epitope) or noncontiguous amino acids juxtaposed by tertiary folding of a protein (conformational epitopes). Epitopes formed from contiguous amino acids are typically retained on exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost on treatment with denaturing solvents.
  • An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. Methods of determining spatial conformation of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed (1996), which is incorporated herein by reference in its entirety.
  • epitope can comprise both dominant and subdominant epitopes.
  • dominant epitope refers to an epitope (e.g., of a tumor antigen) that evokes a strong immune response.
  • substitutedominant epitope refers to an epitope (e.g., of a tumor antigen) that evokes a weak or no immune response.
  • the term “vaccine” refers to an agent that is capable of inducing an immune in a subject upon administration.
  • the vaccine is a "preventive” vaccine, which is administered to a subject not afflicted with a disease or disorder disclosed herein (e.g., cancer). Such vaccines are also referred to herein as “prophylactic” vaccines.
  • the vaccine is a therapeutic vaccine.
  • therapeutic vaccine refers to a vaccine that is administered to a subject to treat a disease or disorder (e.g., prevent or reduce one or more symptoms associated with the disease or disorder).
  • nucleotide vaccine As used herein, the terms “nucleotide vaccine,” “nucleic acid vaccine,” “nucleic acid-based vaccine,” and “genetic vaccine” can be used interchangeably and refer to a vaccine in which the antigenic component comprises a nucleic acid. Such vaccines are capable of delivering genetic materials encoding the antigen of interest (e.g., tumor antigen) into host cells, which subsequently produce the antigen and thereby, initiate an immune response that is capable of protecting the host against the disease or disorder from which the antigen was derived.
  • a nucleotide vaccine comprises both DNA vaccine and RNA (e.g., mRNA) vaccine.
  • a nucleotide vaccine is a DNA vaccine (i.e., the antigenic component is a DNA sequence). In certain aspects, a nucleotide vaccine is a RNA (mRNA) vaccine (i.e., the antigenic component is a RNA sequence).
  • mRNA RNA
  • naturally-occurring refers to the fact that an object can be found in nature.
  • a polypeptide or polynucleotide sequence that is present in an organism (including viruses) that can be isolated from a source in nature and which has not been intentionally modified by man in the laboratory is naturally- occurring.
  • a "polypeptide” refers to a chain comprising at least two consecutively linked amino acid residues, with no upper limit on the length of the chain.
  • One or more amino acid residues in the protein can contain a modification such as, but not limited to, glycosylation, phosphorylation or disulfide bond formation.
  • a “protein” can comprise one or more polypeptides. Unless otherwise specified, the terms “protein” and “polypeptide” can be used interchangeably.
  • the term "nucleic acid molecule,” as used herein, is intended to include DNA molecules and RNA molecules.
  • a nucleic acid molecule can be single- stranded or doublestranded, and can be cDNA.
  • the nucleic acids can be present in whole cells, in a cell lysate, or in a partially purified or substantially pure form.
  • a nucleic acid is "isolated” or “rendered substantially pure” when purified away from other cellular components or other contaminants, e.g., other cellular nucleic acids (e.g., the other parts of the chromosome) or proteins, by standard techniques, including alkaline/SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis and others well known in the art. See, F. Ausubel, et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).
  • Nucleic acids e.g., cDNA
  • cDNA can be mutated, in accordance with standard techniques to provide gene sequences. For coding sequences, these mutations, can affect amino acid sequence as desired.
  • DNA sequences substantially homologous to or derived from native V, D, J, constant, switches and other such sequences described herein are contemplated (where "derived" indicates that a sequence is identical or modified from another sequence).
  • Constant amino acid substitutions refer to substitutions of an amino acid residue with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).
  • basic side chains e.
  • a predicted nonessential amino acid residue in an antibody is replaced with another amino acid residue from the same side chain family.
  • Methods of identifying nucleotide and amino acid conservative substitutions which do not eliminate antigen binding are well-known in the art (see, e.g., Brummell et al., Biochem. 32: 1180-1187 (1993); Kobayashi et al. Protein Eng. 12(10):879-884 (1999); and Burks et al. Proc. Natl. Acad. Sci. USA 94:412-417 (1997)).
  • nucleic acids For nucleic acids, the term “substantial homology” indicates that two nucleic acids, or designated sequences thereof, when optimally aligned and compared, are identical, with appropriate nucleotide insertions or deletions, in at least about 80% of the nucleotides, at least about 90% to 95%, or at least about 98% to 99.5% of the nucleotides. Alternatively, substantial homology exists when the segments will hybridize under selective hybridization conditions, to the complement of the strand.
  • polypeptides the term “substantial homology” indicates that two polypeptides, or designated sequences thereof, when optimally aligned and compared, are identical, with appropriate amino acid insertions or deletions, in at least about 80% of the amino acids, at least about 90% to 95%, or at least about 98% to 99.5% of the amino acids.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, e.g., as described in the non-limiting examples below.
  • the percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at worldwideweb.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6.
  • the percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4: 11-17 (1989)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at worldwideweb.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.
  • nucleic acid and protein sequences described herein can further be used as a "query sequence" to perform a search against public databases to, for example, identify related sequences.
  • Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10.
  • Gapped BLAST can be utilized as described in Altschul et al.. (1997) Nucleic Acids Res. 25(17):3389-3402.
  • the default parameters of the respective programs e.g., XBLAST and NBLAST
  • XBLAST and NBLAST can be used. See worldwideweb.ncbi.nlm.nih.gov.
  • effector function refers to a specialized function of a differentiated immune cell.
  • An effector function of a T cell for example, can be cytolytic activity or helper activity including the secretion of cytokines.
  • An effector function in a naive, memory, or memory-type T cell can also include antigen-dependent proliferation.
  • immune cell refers to cells that play a role in the immune response.
  • immune cells useful for the present disclosure are those cells that can play a role in the treatment and/or eradication of a solid tumor (e.g., possess anti-tumor activity).
  • the immune cells comprise lymphocytes, neutrophils, monocytes, macrophages, dendritic cells, or any combination thereof.
  • the lymphocytes comprise T cells, tumor-infiltrating lymphocytes (TIL), lymphokine- activated killer cells, natural killer T (NKT) cells, or any combination thereof.
  • TIL tumor-infiltrating lymphocytes
  • NKT natural killer T cells
  • the lymphocytes are T cells.
  • the lymphocytes are NKT cells (e.g., invariant NKT cells).
  • vector is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
  • plasmid refers to a circular double stranded DNA loop into which additional DNA segments can be ligated.
  • viral vector Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome.
  • Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors).
  • vectors e.g., non-episomal mammalian vectors
  • vectors can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome.
  • certain vectors are capable of directing the expression of genes to which they are operatively linked.
  • Such vectors are referred to herein as "recombinant expression vectors" (or simply, "expression vectors")
  • expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
  • plasmid and vector can be used interchangeably as the plasmid is the most commonly used form of vector.
  • viral vectors e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses
  • recombinant host cell (or simply “host cell”), as used herein, is intended to refer to a cell that comprises a nucleic acid that is not naturally present in the cell, and can be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell. Because certain modifications can occur in succeeding generations due to either mutation or environmental influences, such progeny cannot, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell” as used herein.
  • a "cancer” refers a broad group of various diseases characterized by the uncontrolled growth of abnormal cells (or tumors) in the body. Unregulated cell division and growth results in the formation of malignant tumors that invade neighboring tissues and can also metastasize to distant parts of the body through the lymphatic system or bloodstream. Cancers that can be treated with the present disclosure include those associated with a solid tumor. Unless indicated otherwise, the terms “cancers” and “tumors” can be used interchangeably.
  • fusion protein refers to proteins created through the joining of two or more genes that originally coded for separate proteins. Translation of this fusion gene results in a single polypeptide or multiple polypeptides with functional properties derived from each of the original proteins.
  • the two or more genes can comprise a substitution, a deletion, and / or an addition in its nucleotide sequence.
  • Fc receptor or “FcR” is a receptor that binds to the Fc region of an immunoglobulin.
  • FcRs that bind to an IgG antibody comprise receptors of the FcyR family, including allelic variants and alternatively spliced forms of these receptors.
  • the FcyR family consists of three activating (FcyRI, FcyRIII, and FcyRIV in mice; FcyRIA, FcyRIIA, and FcyRIIIA in humans) and one inhibitory (FcyRIIB) receptor.
  • FcyRIIB inhibitory receptor
  • NK cells selectively express one activating Fc receptor (FcyRIII in mice and FcyRIIIA in humans) but not the inhibitory FcyRIIB in mice and humans.
  • Human IgGl binds to most human Fc receptors and is considered equivalent to murine IgG2a with respect to the types of activating Fc receptors that it binds to.
  • an "Fc region” fragment crystallizable region or “Fc domain” or “Fc” refers to the C-terminal region of the heavy chain of an antibody that mediates the binding of the immunoglobulin to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells) or to the first component (Clq) of the classical complement system.
  • an Fc region comprises the constant region of an antibody excluding the first constant region immunoglobulin domain (e.g., CHI or CL).
  • the Fc region comprises two identical protein fragments, derived from the second (CH2) and third (CH3) constant domains of the antibody's two heavy chains; IgM and IgE Fc regions comprise three heavy chain constant domains (CH domains 2-4) in each polypeptide chain.
  • the Fc region comprises immunoglobulin domains CH2 and CH3 and the hinge between CHI and CH2 domains.
  • the human IgG heavy chain Fc region is defined to stretch from an amino acid residue D221 for IgGl, V222 for IgG2, L221 for IgG3 and P224 for IgG4 to the carboxy -terminus of the heavy chain, wherein the numbering is according to the EU index as in Kabat.
  • the CH2 domain of a human IgG Fc region extends from amino acid 237 to amino acid 340, and the CH3 domain is positioned on C-terminal side of a CH2 domain in an Fc region, /. ⁇ ., it extends from amino acid 341 to amino acid 447 or 446 (if the C-terminal lysine residue is absent) or 445 (if the C- terminal glycine and lysine residues are absent) of an IgG.
  • the Fc region can be a native sequence Fc, including any allotypic variant, or a variant Fc (e.g., a non-naturally occurring Fc).
  • Fc can also refer to this region in isolation or in the context of an Fc-comprising protein polypeptide such as a "binding protein comprising an Fc region,” also referred to as an “Fc fusion protein” (e.g., an antibody or immunoadhesion).
  • a binding protein comprising an Fc region also referred to as an “Fc fusion protein” (e.g., an antibody or immunoadhesion).
  • a "native sequence Fc region” or “native sequence Fc” comprises an amino acid sequence that is identical to the amino acid sequence of an Fc region found in nature.
  • Native sequence human Fc regions include a native sequence human IgGl Fc region; native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region as well as naturally occurring variants thereof.
  • Native sequence Fc include the various allotypes of Fes (see, e.g., Jefferis et al. (2009) mAbs 1 : 1).
  • an Fc (native or variant) of the present disclosure can be in the form of having native sugar chains, increased sugar chains, or decreased sugar chains compared to the native form, or can be in a deglycosylated form.
  • the immunoglobulin Fc sugar chains can be modified by conventional methods such as a chemical method, an enzymatic method, and a genetic engineering method using a microorganism. The removal of sugar chains from an Fc fragment results in a sharp decrease in binding affinity to the Clq part of the first complement component Cl, and a decrease or loss of ADCC or CDC, thereby not inducing any unnecessary immune responses in vivo.
  • an immunoglobulin Fc region in a deglycosylated or aglycosylated form can be more suitable to the object of the present disclosure as a drug carrier.
  • deglycosylation refers to an Fc region in which sugars are removed enzymatically from an Fc fragment.
  • aglycosylation means that an Fc fragment is produced in an unglycosylated form by a prokaryote, and preferably in E. coli.
  • an immune response refers to a biological response within a vertebrate against foreign agents, which response protects the organism against these agents and diseases caused by them.
  • An immune response is mediated by the action of a cell of the immune system (e.g., a T lymphocyte, B lymphocyte, natural killer (NK) cell, macrophage, eosinophil, mast cell, dendritic cell or neutrophil) and soluble macromolecules produced by any of these cells or the liver (including antibodies, cytokines, and complement) that results in selective targeting, binding to, damage to, destruction of, and/or elimination from the vertebrate's body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues.
  • An immune reaction includes, e.g., activation or inhibition of a T cell, e.g., an effector T cell or a Th cell, such as
  • an immune response (e.g., such as that induced by a nucleotide vaccine disclosed herein) comprises a T cell immune response.
  • T cell immune response refers to an immune response mediated by T cells (e.g., effector CD4 + and/or CD8 + T cells).
  • a T cell immune response can be generally divided into three phases: (i) expansion, (ii) contraction, and (iii) maintenance. Kumar et al., Immunity 48(2): 202-213 (Feb. 2018); and Blair et al., J Immunol 187: 2310-2321 (2011).
  • naive T cells that recognize their cognate antigen become activated, resulting in clonal expansion and acquisition of effector function (e.g., production of inflammatory cytokines and expression of effector molecules, such as granzyme and perforin).
  • effector function e.g., production of inflammatory cytokines and expression of effector molecules, such as granzyme and perforin.
  • approximately 90-95% of the activated T cells at the peak of the response undergo apoptosis (i.e., contraction phase).
  • the surviving population of activated T cells eventually differentiate into memory T cells that provide long-lasting protection to the host (i.e., maintenance phase).
  • immunotherapy refers to the treatment of a subject afflicted with, or at risk of contracting or suffering a recurrence of, a disease by a method comprising inducing, enhancing, suppressing or otherwise modifying an immune response.
  • Treatment or “therapy” of a subject refers to any type of intervention or process performed on, or the administration of an active agent to, the subject with the objective of reversing, alleviating, ameliorating, inhibiting, slowing down or preventing the onset, progression, development, severity or recurrence of a symptom, complication or condition, or biochemical indicia associated with a disease.
  • tumor infiltrating lymphocytes refers to lymphocytes (e.g., effector T cells) that have migrated from the periphery (e.g., from the blood) into a tumor.
  • the tumor infiltrating lymphocytes are CD4+ TILs.
  • the tumor infiltrating lymphocytes are CD8+ TILs.
  • An increased ability to stimulate an immune response or the immune system can result from an enhanced agonist activity of T cell costimulatory receptors and/or an enhanced antagonist activity of inhibitory receptors.
  • An increased ability to stimulate an immune response or the immune system can be reflected by a fold increase of the EC50 or maximal level of activity in an assay that measures an immune response, e.g., an assay that measures changes in cytokine or chemokine release, cytolytic activity (determined directly on target cells or indirectly via detecting CD 107a or granzymes) and proliferation.
  • the ability to stimulate an immune response or the immune system activity can be enhanced by at least 10%, 30%, 50%, 75%, 2 fold, 3 fold, 5 fold or more.
  • interleukin-7 refers to IL-7 polypeptides and derivatives and analogs thereof having substantial amino acid sequence identity to wild-type mature mammalian IL-7 proteins and substantially equivalent biological activity, e.g, in standard bioassays or assays of IL-7 receptor binding affinity. Additional disclosure relating to IL-7 proteins that can be used with the present disclosure are provided elsewhere herein.
  • a "variant” of an IL-7 protein is defined as an amino acid sequence that is altered by one or more amino acids.
  • the variant can have "conservative” changes, wherein a substituted amino acid has similar structural or chemical properties, e.g, replacement of leucine with isoleucine. More rarely, a variant can have "nonconservative” changes, e.g., replacement of a glycine with a tryptophan. Similar minor variations can also include amino acid deletions or insertions, or both.
  • Guidance in determining which and how many amino acid residues can be substituted, inserted or deleted without abolishing biological activity can be found using computer programs well known in the art, for example software for molecular modeling or for producing alignments.
  • the variant IL-7 proteins included within the present disclosure include IL-7 proteins that retain IL-7 activity.
  • IL-7 polypeptides which also include additions, substitutions or deletions are also included within the present disclosure as long as the proteins retain substantially equivalent biological IL-7 activity.
  • truncations of IL-7 which retain comparable biological activity as the full length form of the IL-7 protein are included within the present disclosure.
  • variant IL-7 proteins also include polypeptides that have at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more sequence identity with wild-type IL-7.
  • signal sequence refers to a fragment directing the secretion of a biologically active molecule drug and a fusion protein, and it is cut off after being translated in a host cell.
  • the signal sequence as used herein is a polynucleotide encoding an amino acid sequence initiating the movement of the protein penetrating the endoplasmic reticulum (ER) membrane.
  • useful signal sequences include an antibody light chain signal sequence, e.g., antibody 14.18 (Gillies et al.., J. Immunol. Meth 1989.
  • an antibody heavy chain signal sequence e.g., MOPC141 an antibody heavy chain signal sequence (Sakano et al., Nature, 1980.286: 676-683), and other signal sequences know in the art (e.g., see Watson et al., Nucleic Acid Research, 1984.12:5145-5164).
  • the characteristics of signal peptides are well known in the art, and the signal peptides conventionally having 16 to 30 amino acids, but they can include more or less number of amino acid residues.
  • Conventional signal peptides consist of three regions of the basic N-terminal region, a central hydrophobic region, and a more polar C-terminal region.
  • a “subject” includes any human or nonhuman animal.
  • the term “nonhuman animal” includes, but is not limited to, vertebrates such as nonhuman primates, sheep, dogs, and rodents such as mice, rats and guinea pigs. In some aspects, the subject is a human.
  • the terms “subject” and “patient” are used interchangeably herein.
  • the term “therapeutically effective amount” or “therapeutically effective dosage” refers to an amount of an agent that provides the desired biological, therapeutic, and/or prophylactic result. That result can be reduction, amelioration, palliation, lessening, delaying, and/or alleviation of one or more of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system.
  • an effective amount comprises an amount sufficient to cause a tumor to shrink and/or to decrease the growth rate of the tumor (such as to suppress tumor growth) or to prevent or delay other unwanted cell proliferation. In some aspects, an effective amount is an amount sufficient to delay tumor development. In some aspects, an effective amount is an amount sufficient to prevent or delay tumor recurrence. An effective amount can be administered in one or more administrations.
  • the effective amount of the drug or composition can: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, retard, slow to some extent and can stop cancer cell infiltration into peripheral organs; (iv) inhibit (i.e., slow to some extent and can stop tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay occurrence and/or recurrence of tumor; and/or (vii) relieve to some extent one or more of the symptoms associated with the cancer.
  • a "therapeutically effective amount” is the amount of nucleotide vaccine and/or IL-7 protein clinically proven to affect a significant decrease in cancer or slowing of progression (regression) of cancer, such as an advanced solid tumor.
  • the ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays.
  • dosing frequency/ dosing schedule and “dosing interval” are used interchangeably and refer to the number of times a therapeutic agent (e.g., a nucleotide vaccine and/or IL-7) is administered to a subject within a specific time period. Dosing frequency can be indicated as the number of doses per a given time, for example, once per day, once a week, or once in two weeks. As used herein, “dosing frequency” is applicable where a subject receives multiple (or repeated) administrations of a therapeutic agent.
  • a therapeutic agent e.g., a nucleotide vaccine and/or IL-7
  • the term "within” when used to describe an administration of a therapeutic agent described herein means that the therapeutic agent is administered on or before the accompanying duration of time.
  • a therapeutic agent described herein e.g., a nucleotide vaccine and/or IL-7
  • standard of care refers to a treatment that is accepted by medical experts as a proper treatment for a certain type of disease and that is widely used by healthcare professionals.
  • the term can be used interchangeable with any of the following terms: “best practice,” “standard medical care,” and “standard therapy.”
  • an "anti-cancer agent” promotes cancer regression in a subject or prevents further tumor growth.
  • a therapeutically effective amount of the drug promotes cancer regression to the point of eliminating the cancer.
  • Promote cancer regression means that administering an effective amount of the drug, alone or in combination with an anti-neoplastic agent, results in a reduction in tumor growth or size, necrosis of the tumor, a decrease in severity of at least one disease symptom, an increase in frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction.
  • the terms "effective” and “effectiveness” with regard to a treatment includes both pharmacological effectiveness and physiological safety.
  • Pharmacological effectiveness refers to the ability of the drug to promote cancer regression in the patient.
  • Physiological safety refers to the level of toxicity, or other adverse physiological effects at the cellular, organ and/or organism level (adverse effects) resulting from administration of the drug.
  • a therapeutically effective amount of an anti-cancer agent can inhibit cell growth or tumor growth by at least about 10%, at least about 20%, by at least about 40%, by at least about 60%, or by at least about 80% relative to untreated subjects or, in certain aspects, relative to patients treated with a standard-of-care therapy.
  • tumor regression can be observed and continue for a period of at least about 20 days, at least about 40 days, or at least about 60 days. Notwithstanding these ultimate measurements of therapeutic effectiveness, evaluation of immunotherapeutic drugs must also make allowance for "immune-related" response patterns.
  • immune checkpoint inhibitor refers to molecules that totally or partially reduce, inhibit, interfere with or modulate one or more immune checkpoint proteins.
  • Immune checkpoint proteins regulate T-cell activation or function. Numerous checkpoint proteins are known, such as CTLA-4 and its ligands CD80 and CD86; and PD-1 with its ligands PD-L1 and PD-L2. Pardoll, D.M., Nat Rev Cancer 12(4):252-64 (2012). These proteins are responsible for co-stimulatory or inhibitory interactions of T-cell responses. Immune checkpoint proteins regulate and maintain self-tolerance and the duration and amplitude of physiological immune responses. Immune checkpoint inhibitors include antibodies or are derived from antibodies.
  • a method for treating a tumor (or a cancer associated with a tumor) in a subject in need thereof comprising administering to the subject a nucleotide vaccine in combination with an interleukin-7 (IL-7) protein.
  • the nucleotide vaccine encodes a tumor antigen, such that the administration of the nucleotide vaccine is capable of inducing a tumor-specific T cell immune response in the subject.
  • the IL-7 protein is administered to the subject after the peak expansion phase of the tumorspecific T cell immune response induced by the nucleotide vaccine.
  • the term "peak expansion phase” refers to the time point at which the number of tumor-specific T cells, e.g., induced by the nucleotide vaccine, is the greatest. In some aspects, the peak expansion phase marks the beginning of the contraction phase of a T cell immune response. As is apparent from the present disclosure, the peak expansion phase of the nucleotide vaccine-induced tumorspecific T cell immune response can differ depending on whether the nucleotide vaccine is administered therapeutically (z.e., after the occurrence of a tumor) or prophylactically (/. ⁇ ., before the occurrence of a tumor). The peak expansion phase of a T cell immune response can be determined using any suitable methods known in the art (e.g., ELISPOT and flow cytometry).
  • the peak expansion of the nucleotide vaccine-induced tumor-specific T cell immune response occurs at about seven days, about eight days, about nine days, about ten days, about 11 days, about 12 days, about 13 days, about 14 days, or about 15 days after initial activation (z.e., antigen encounter) of the antigen-specific T cells (e.g., after initial administration of a nucleotide vaccine encoding a tumor antigen to the subject).
  • the peak expansion of the tumor-specific T cell immune response occurs at about 11 days after initial activation of the antigen-specific T cells (e.g., after initial administration of a nucleotide vaccine encoding a tumor antigen to the subject).
  • the peak expansion can occur earlier.
  • tumor-specific T cells can already exist in the subject from an earlier encounter with the existing tumor.
  • the existing tumor-specific T cells e.g., memory T cells
  • the existing tumor-specific T cells could respond more quickly (compared to T cells that are seeing the antigen for the first time), resulting in a faster T cell kinetics (z.e., an earlier peak expansion).
  • an IL-7 protein after the peak expansion phase can improve a tumor-specific T cell immune response (e.g., induced by a nucleotide vaccine disclosed herein) compared to the corresponding value (z.e., tumor-specific T cell immune response) observed in a reference.
  • a tumor-specific T cell immune response e.g., induced by a nucleotide vaccine disclosed herein
  • corresponding value z.e., tumor-specific T cell immune response
  • the term "reference" can refer to a corresponding individual that (i) only received the nucleotide vaccine alone, (ii) only received the IL-7 protein alone, (iii) received both the nucleotide vaccine and IL-7 protein, but the IL-7 protein was administered prior to the peak expansion phase, (iv) received neither the nucleotide vaccine nor the IL-7 protein, or (iv) combinations thereof.
  • an IL-7 protein described herein is generally administered to the subject after the administration of the nucleotide vaccine.
  • Applicant has further identified that administering an IL-7 protein at other time points (e.g. , other than after the peak expansion phase of the tumor-specific T cell immune response) after nucleotide vaccine administration can also have therapeutic effects, e.g, particularly where the nucleotide vaccine is administered as a therapeutic vaccine.
  • the IL-7 protein is administered to the subject within about six hours, within about 12 hours, within about one day, within about two days, within about three days, within about four days, within about five days, within about six days, within about one week, within about two weeks, without about three weeks, or within about four weeks after the administration of the nucleotide vaccine administration.
  • the IL-7 protein is administered within about one week after the nucleotide vaccine administration.
  • the IL-7 protein is administered within about five days after the nucleotide vaccine administration.
  • the IL-7 protein is administered within about four days after the nucleotide vaccine administration.
  • the IL-7 protein is administered within about three days after the nucleotide vaccine administration.
  • the IL-7 protein is administered within about two days after the nucleotide vaccine administration. In some aspects, the IL-protein is administered within about one day after the nucleotide vaccine administration. In some aspects, the IL-7 protein is administered concurrently with the nucleotide vaccine administration.
  • an improved tumor-specific T cell immune response can result in reduced tumor growth in the subject.
  • administering a nucleotide vaccine in combination with an IL-7 protein, wherein the IL-7 protein is administered after the peak expansion phase of the tumor-specific T cell immune response can inhibit and/or reduce tumor growth (e.g., tumor volume or weight) in the subject compared to the reference.
  • the tumor growth is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% after the administration compared to the corresponding value (i.e., tumor growth) in the reference.
  • administering an IL-7 protein i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine can inhibit and/or reduce tumor growth (e.g., tumor volume or weight) in the subject compared to the reference.
  • the tumor growth is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% after the administration compared to the corresponding value (i.e., tumor growth) in the reference.
  • an improved tumor-specific T cell immune response can result in a more prolonged tumor-specific T cell immune response in the subject.
  • the prolonged tumor-specific T cell immune response is due to increased survival (e.g., during the contraction phase) of the tumor-specific T cells.
  • administering a nucleotide vaccine in combination with an IL-7 protein, wherein the IL-7 protein is administered after the peak expansion phase of the tumorspecific T cell immune response can increase the survival of the tumor-specific T cells (e.g., during the contraction phase) in the subject compared to the corresponding value in the reference.
  • the survival of the tumor-specific T cells is increased by at least about 1-fold, at least about 2-fold, at least about 3- fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the corresponding value in the reference.
  • administering an IL-7 protein (i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine can prolong the tumor-specific T cell immune response in the subject compared to the reference.
  • the tumor-specific T cell immune response is prolonged by at least about 1-fold, at least about 2-fold, at least about 3 -fold, at least about 4- fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the corresponding value in the reference.
  • the prolonged tumor-specific T cell immune response is due to increased resistance of the tumor-specific T cells to apoptosis (e.g., during the contraction phase of the immune response).
  • the resistance of the tumor-specific T cells to apoptosis is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6- fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the corresponding value in the reference.
  • increasing the initial expansion of the tumor-specific T cells can also help prolong a tumor-specific T cell immune response (e.g., by increasing the overall number of tumor-specific T cells).
  • administering an IL-7 protein can increase the initial expansion of the nucleotide vaccine-induced tumor-specific T cells by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more compared to the corresponding value in the reference.
  • an increased survival and/or increased resistance to apoptosis can increase the number of tumor-specific T cells in the subject compared to the corresponding value in the reference.
  • the number of tumor-specific T cells in the subject is increased by at least about 1-fold, at least about 2-fold, at least about 3 -fold, at least about 4- fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the corresponding value in the reference.
  • an improved tumor-specific T cell immune response comprises an increased cytotoxic activity of the tumor-specific T cells.
  • administering a nucleotide vaccine in combination with an IL-7 protein increases the ability of the tumor-specific T cells to kill a cell expressing a tumor antigen (e.g., tumor cell).
  • the cytotoxic activity of the tumor-specific T cells is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10- fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the corresponding value in the reference (e.g., corresponding subject that did not receive the combination treatment).
  • an expansion phase of a T cell immune response is generally followed by a contraction phase, during which a large fraction (e.g., 90-95%) of the activated effector T cells undergo apoptosis with the surviving effector T cells differentiating into long- lived memory T cells.
  • a large fraction e.g. 90-95%
  • the increased survival and/or resistance of the tumor-specific T cells (e.g, during the contraction phase) to apoptosis can result in greater number of tumor-specific memory T cells in the subject.
  • increasing the initial expansion of the tumor-specific T cells can also help result in greater number of tumor-specific memory T cells in the subject.
  • the number of tumorspecific memory T cells in the subject is increased by at least about 1-fold, at least about 2- fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the corresponding value in the reference.
  • an improved tumor-specific T cell immune response comprises an expanded T cell receptor repertoire of a tumor-specific T cell immune response.
  • methods of the present disclosure e.g., administering a nucleotide vaccine in combination with IL-7, wherein the IL-7 is administered to subject after the peak expansion phase of a tumor-specific T cell immune response
  • administering an IL-7 protein (i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine can increase the number of epitopes against which the tumor-specific T cell immune response is induced, compared to the corresponding value in the reference.
  • the number of epitopes against which the tumor-specific T cell immune response is induced is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7- fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to the reference.
  • tumor-specific T cells disclosed herein (/. ⁇ ., induced after administration of a nucleotide vaccine encoding a tumor antigen in combination with IL-7, wherein the IL-7 is administered after the peak expansion phase of a tumor-specific T cell immune response) is capable of targeting one or more epitopes of a tumor antigen comprising Lrrc27, Plekhol, Pttgl, Xpo4, Exoc4, Pank3, TmemlOl, Map3k6, Met, BC057079, Histlh3e, Prkagl, Neil3, or combinations thereof.
  • tumor-specific T cells produced after the administration of an IL-7 protein can target one or more epitopes of a tumor antigen comprising Lrrc27, Plekhol, Pttgl, Xpo4, Exoc4, Pank3, TmemlOl, Map3k6, Met, BC057079, Histlh3e, Prkagl, Neil3, or combinations thereof. Additional disclosures regarding tumor antigens that can be targeted using the methods disclosed herein are provided else wherein the present disclosure.
  • tumorspecific T cells described herein can target one, two, three, four, five, six, seven, eight, nine, ten, 11, 12, or all of the tumor epitopes described above.
  • the tumor-specific T cells of the present disclosure are capable of targeting the following epitopes: Lrrc27, Plekhol, and Pttgl.
  • the present disclosure is directed to a method of increasing a T cell immune response against a subdominant epitope of a tumor antigen in a subject in need thereof, comprising administering to the subject a nucleotide vaccine encoding a tumor antigen, which comprises the subdominant epitope, in combination with an IL-7, wherein the IL-7 is administered to the subject after the peak expansion phase of the tumorspecific T cell immune response.
  • a method of increasing a T cell immune response against a subdominant epitope of a tumor antigen in a subject in need thereof comprises administering to the subject a nucleotide vaccine encoding a tumor antigen, which comprises the subdominant epitope, in combination with an IL-7, wherein the IL-7 protein is administered: (i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine.
  • a T cell immune response against a subdominant epitope of a tumor antigen is increased by at least about 1-fold, at least about 2- fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold or more, compared to a corresponding value in a reference.
  • methods of the present disclosure comprises administering a nucleotide vaccine encoding a tumor antigen in combination with IL-7, wherein the IL-7 is administered after the peak expansion phase of a tumor-specific T cell immune response.
  • the IL-7 is administered to the subject (e.g., suffering from a tumor) at least about one day, about two days, about three days, about four days, about five days, about six days, about seven days, about eight days, about nine days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, or about 30 days or more after the peak expansion phase of the tumor-specific T cell immune response.
  • the IL-7 is administered at about two days after the peak expansion phase of the tumor-specific T cell immune response.
  • Non-limiting examples of cancers (or tumors) that can be treated with methods disclosed herein include squamous cell carcinoma, small-cell lung cancer (SCLC), non-small cell lung cancer, squamous non-small cell lung cancer (NSCLC), nonsquamous NSCLC, gastrointestinal cancer, renal cancer (e.g., clear cell carcinoma), ovarian cancer, liver cancer (e.g., hepatocellular carcinoma), colorectal cancer, endometrial cancer, kidney cancer (e.g., renal cell carcinoma (RCC)), prostate cancer (e.g., hormone refractory prostate adenocarcinoma), thyroid cancer, pancreatic cancer, cervical cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon carcinoma, and head and neck cancer (or carcinoma), gastric cancer, germ cell tumor, pediatric sarcoma, sinonasal natural killer, melanoma (e.g., metastatic malignant melanoma, such as cutaneous or intraocular malignant melanom
  • a cancer (or tumor) that can be treated comprises a breast cancer, head and neck cancer, uterine cancer, brain cancer, skin cancer, renal cancer, lung cancer, colorectal cancer, prostate cancer, liver cancer, bladder cancer, kidney cancer, pancreatic cancer, thyroid cancer, esophageal cancer, eye cancer, stomach (gastric) cancer, gastrointestinal cancer, carcinoma, sarcoma, leukemia, lymphoma, myeloma, or a combination thereof.
  • a cancer (or tumor) that can be treated with the present methods is breast cancer.
  • breast cancer is a triple negative breast cancer (TNBC).
  • a cancer (or tumor) that can be treated is a brain cancer.
  • brain cancer is a glioblastoma.
  • a cancer (or tumor) that can be treated with the present methods is skin cancer.
  • skin cancer is a basal cell carcinoma (BCC), cutaneous squamous cell carcinoma (cSCC), melanoma, Merkel cell carcinoma (MCC), or a combination thereof.
  • a head and neck cancer is a head and neck squamous cell carcinoma.
  • a lung cancer is a small cell lung cancer (SCLC).
  • SCLC small cell lung cancer
  • an esophageal cancer is gastroesophageal junction cancer.
  • a kidney cancer is renal cell carcinoma.
  • a liver cancer is hepatocellular carcinoma.
  • the methods described herein can also be used for treatment of metastatic cancers, unresectable, refractory cancers (e.g., cancers refractory to previous cancer therapy, e.g., immunotherapy, e.g., with a blocking anti-PD-1 antibody), and/or recurrent cancers.
  • metastatic cancers unresectable, refractory cancers (e.g., cancers refractory to previous cancer therapy, e.g., immunotherapy, e.g., with a blocking anti-PD-1 antibody), and/or recurrent cancers.
  • the previous cancer therapy comprises a chemotherapy.
  • the chemotherapy comprises a platinum-based therapy.
  • the platinum-based therapy comprises a platinum-based antineoplastic selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, and any combination thereof.
  • the platinum-based therapy comprises cisplatin.
  • the platinum-based therapy comprises carboplatin.
  • methods disclosed herein e.g., administering a nucleotide vaccine encoding a tumor antigen in combination with IL-7, wherein the IL-7 is administered after the peak expansion phase of a tumor-specific T cell immune response or wherein the IL-7 is administered: (i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine) effectively increases the duration of survival of a subject in need thereof (e.g., afflicted with a tumor).
  • duration of survival of the subject is increased by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least about 1 year or more when compared to a reference individual (e.g., corresponding subject treated with IL-7 protein alone or with a bispecific antibody alone).
  • a reference individual e.g., corresponding subject treated with IL-7 protein alone or with a bispecific antibody alone.
  • the methods disclosed herein increases duration of survival of the subject at a level higher than (about one month higher than, about two months higher than, about three months higher than, about four months higher than, about five months higher than, about six months higher than, about seven months higher than, about eight months higher than, about nine months higher than, about ten months higher than, about eleven months higher than, or about one year higher than) the duration of survival of a reference subject.
  • methods of the present disclosure e.g., administering a nucleotide vaccine encoding a tumor antigen in combination with IL-7, wherein the IL-7 is administered after the peak expansion phase of a tumor-specific T cell immune response or wherein the IL- 7 is administered: (i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine) effectively increase the duration of progression- free survival of a subject (e.g., cancer patient).
  • a subject e.g., cancer patient
  • the progression free survival of the subject is increased by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least about 1 year when compared to a reference subject.
  • methods disclosed herein e.g., administering a nucleotide vaccine encoding a tumor antigen in combination with IL-7, wherein the IL-7 is administered after the peak expansion phase of a tumor-specific T cell immune response or wherein the IL-7 is administered: (i) after the administration of the nucleotide vaccine or (ii) concurrently with the administration of the nucleotide vaccine) effectively increases the response rate in a group of subjects.
  • the response rate in a group of subjects is increased by at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at last about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99% or at least about 100% when compared to a reference subject.
  • the combination therapy described herein e.g., combination of a nucleotide vaccine and an IL-7 protein
  • a prophylactic treatment against a disease or disorder described herein e.g., cancer
  • a method of preventing or reducing the occurrence of a tumor in a subject in need thereof comprising administering to the subject a nucleotide vaccine encoding a tumor antigen in combination with an IL-7 protein, wherein the administration of the nucleotide vaccine induces a tumor-specific T cell immune response, and wherein the nucleotide vaccine, the IL-7 protein, or both the nucleotide vaccine and the IL-7 protein are administered to the subject prior to the occurrence of the tumor.
  • the nucleotide vaccine, the IL-7 protein, or both the nucleotide vaccine and the IL-7 protein are administered to the subject at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about two weeks, at least about three weeks, at least about four weeks, at least about two months, at least about three months, at least about four months, at least about five months, at least about six months, at least about seven months, at least about eight months, at least about nine months, at least about 10 months, at least 11 months, or at least about one year prior to the occurrence of a tumor.
  • administering the nucleotide vaccine, the IL-7 protein, or both the nucleotide vaccine and the IL-7 protein as described above can help reduce the occurrence of the tumor by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70% at least about 80%, at least about 90%, or at least about 100%, compared to a reference (e.g., corresponding subject that did not receive the combination treatment as described above).
  • a reference e.g., corresponding subject that did not receive the combination treatment as described above.
  • the subject being treated in the methods of the present disclosure is a nonhuman animal, such as a rat or a mouse. In some aspects, the subject being treated is a human.
  • IL-7 (e.g., such as those disclosed herein) is administered at a weight-based dose. In certain aspects, the IL-7 is administered at a dose between about 5 mg/kg and about 15 mg/kg. In some aspects, the IL-7 is administered at a dose of about 5 mg/kg.
  • a nucleotide vaccine described herein e.g., encoding a tumor antigen and/or IL-7) can be administered at a dosage in the range of about 0.1 pg to about 200 mg. In certain aspects, the dosage is in the range of about 0.6 mg to about 100 mg. In further aspects, the dosage is in the range of about 1.2 mg to about 50 mg.
  • each dose of a nucleotide vaccine encoding a tumor antigen is about 4 pg.
  • methods disclosed herein comprise administering a single dose of the nucleotide vaccine to a subject (e.g., suffering from a tumor).
  • multiple does of the nucleotide vaccines are administered to the subject.
  • the nucleotide vaccine is administered to the subject at a dosing frequency of about once a day, about once every two days, about once every three days, about once every four days, about once every five days, about once every six days, or about once every seven days.
  • the nucleotide vaccine is administered about once every seven days (i.e., once a week).
  • the nucleotide vaccine is administered about once a month.
  • the nucleotide vaccine is administered about once every three days for a total of three doses.
  • methods disclosed herein comprise administering a single dose of IL-7 to a subject (e.g., suffering from a tumor).
  • the subject receives multiple doses of IL-7 (i.e., repeated administration).
  • the IL-7 can be administered at a dosing frequency of once a week, once in two weeks, once in three weeks, once in four weeks, once in five weeks, once in six weeks, once in seven weeks, once in eight weeks, once in nine weeks, once in 10 weeks, once in 11 weeks, or once in 12 weeks.
  • methods disclosed herein comprise administering to a subject (i) three doses of a nucleotide vaccine (e.g., encoding a tumor antigen) at a dosing frequency of once every three days, and (ii) a single dose of IL-7 after the peak expansion phase of the tumor-specific T cell immune response.
  • the single dose of IL-7 is administered at day 13 post initial administration of the nucleotide vaccine.
  • methods provided herein comprise administering multiple doses of a nucleotide vaccine (e.g., encoding a tumor antigen) in combination with an IL-7 protein, wherein the IL-7 protein is administered after at least one of the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about four weeks after at least one of the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about three weeks after at least one of the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about two weeks after at least one of the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about 13 days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about 12 days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about 11 days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about 10 days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about nine days after at least one of the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about eight days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about one week after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about six days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about five days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about four days after at least one of the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about three days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about two days after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about one day after at least one of the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered to the subject concurrently with at least one of the multiple doses of the nucleotide vaccine.
  • methods provided herein comprise administering to a subject (i) three doses of a nucleotide vaccine e.g., encoding a tumor antigen) at a dosing frequency of once every three days, and (ii) an IL-7 protein, wherein the IL-7 protein is administered within about two days after the administration of at least one of the doses of the nucleotide vaccine.
  • a nucleotide vaccine e.g., encoding a tumor antigen
  • methods provided herein comprise administering to a subject (i) three doses of a nucleotide vaccine (e.g., encoding a tumor antigen) at a dosing frequency of once every three to five days, and (ii) an IL-7 protein, wherein the IL-7 protein is administered within about one day after the administration of at least one of the doses of the nucleotide vaccine.
  • a nucleotide vaccine e.g., encoding a tumor antigen
  • methods provided herein comprise administering to a subject (i) three doses of a nucleotide vaccine (e.g., encoding a tumor antigen) at a dosing frequency of once every three days, and (ii) an IL-7 protein, wherein the IL-7 protein is administered concurrently with at least one of the doses of the nucleotide vaccine.
  • a nucleotide vaccine e.g., encoding a tumor antigen
  • the IL-7 protein is administered to the subject after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about four weeks after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about three weeks after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about two weeks after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about 13 days after administering all the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about 12 days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about 11 days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about 10 days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about nine days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about eight days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about one week after administering all the multiple doses of the nucleotide vaccine.
  • the IL-7 protein is administered within about six days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about five days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about four days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about three days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about two days after administering all the multiple doses of the nucleotide vaccine. In some aspects, the IL-7 protein is administered within about one day after administering all the multiple doses of the nucleotide vaccine.
  • an IL-7 protein disclosed herein can be administered to a subject at a weight-based dose.
  • an IL-7 protein can be administered at a weight-based dose between about 20 pg/kg and about 600 pg/kg.
  • an IL-7 protein of the present disclosure can be administered at a weight-based dose of about 20 pg/kg, about 60 pg/kg, about 120 pg/kg, about 240 pg/kg, about 360 pg/kg, about 480 pg/kg, or about 600 hg/kg-
  • an IL-7 protein disclosed herein can be administered to a subject at a dose greater than about 600 pg/kg.
  • an IL-7 protein is administered to a subject at a dose greater than about 600 pg/kg, greater than about 700 pg/kg, greater than about 800 pg/kg, greater than about 900 pg/kg, greater than about 1,000 pg/kg, greater than about 1,100 pg/kg, greater than about 1,200 pg/kg, greater than about 1,300 pg/kg, greater than about 1,400 pg/kg, greater than about 1,500 pg/kg, greater than about 1,600 pg/kg, greater than about 1,700 pg/kg, greater than about 1,800 pg/kg, greater than about 1,900 pg/kg, or greater than about 2,000 pg/kg.
  • an IL-7 protein of the present disclosure is administered at a dose of between 610 pg/kg and about 1,200 pg/kg, between 650 pg/kg and about 1,200 pg/kg, between about 700 pg/kg and about 1,200 pg/kg, between about 750 pg/kg and about 1,200 pg/kg, between about 800 pg/kg and about 1,200 pg/kg, between about 850 pg/kg and about 1,200 pg/kg, between about 900 pg/kg and about 1,200 pg/kg, between about 950 pg/kg and about 1,200 pg/kg, between about 1,000 pg/kg and about 1,200 pg/kg, between about 1,050 pg/kg and about 1,200 pg/kg, between about 1,100 pg/kg and about 1,200 pg/kg, between about 1,200 pg/kg and about 2,000 pg/kg, between about 1,300 pg
  • an IL-7 protein of the present disclosure is administered at a dose of between 610 pg/kg and about 1,200 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of between 650 pg/kg and about 1,200 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 700 pg/kg and about 1,200 pg/kg. In further aspects, an IL-7 protein is administered at a dose of between about 750 pg/kg and about 1,200 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of between about 800 pg/kg and about 1,200 pg/kg.
  • an IL-7 protein is administered at a dose of between about 850 pg/kg and about 1,200 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 900 pg/kg and about 1,200 pg/kg. In further aspects, an IL-7 protein is administered at a dose of between about 950 pg/kg and about 1,200 pg/kg. In some aspects, an IL-7 protein disclosed herein is administered at a dose of between about 1,000 pg/kg and about 1,200 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 1,050 pg/kg and about 1,200 pg/kg.
  • an IL-7 protein is administered at a dose of between about 1,100 pg/kg and about 1,200 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 1,200 pg/kg and about 2,000 pg/kg. In further aspects, an IL-7 protein is administered at a dose of between about 1,300 pg/kg and about 2,000 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 1,500 pg/kg and about 2,000 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 1,700 pg/kg and about 2,000 pg/kg.
  • an IL-7 protein is administered at a dose of between about 610 pg/kg and about 1,000 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 650 pg/kg and about 1,000 pg/kg. In further aspects, an IL-7 protein is administered at a dose of between about 700 pg/kg and about 1,000 pg/kg. In yet further aspects, an IL-7 protein is administered at a dose of between about 750 pg/kg and about 1,000 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of between about 800 pg/kg and about 1,000 pg/kg.
  • an IL-7 protein is administered at a dose of between about 850 pg/kg and about 1,000 pg/kg. In some aspects, an IL-7 protein of the present disclosure is administered at a dose of between about 900 pg/kg and about 1,000 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 950 pg/kg and about 1,000 pg/kg.
  • an IL-7 protein is administered at a dose of between about 700 pg/kg and about 900 pg/kg, between about 750 pg/kg and about 950 pg/kg, between about 700 pg/kg and about 850 pg/kg, between about 750 pg/kg and about 850 pg/kg, between about 700 pg/kg and about 800 pg/kg, between about 800 pg/kg and about 900 pg/kg, between about 750 pg/kg and about 850 pg/kg, or between about 850 pg/kg and about 950 pg/kg.
  • an IL-7 protein is administered at a dose of between about 700 pg/kg and about 900 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of between about 750 pg/kg and about 950 pg/kg. In further aspects, an IL-7 protein is administered at a dose of between about 700 pg/kg and about 850 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 750 pg/kg and about 850 pg/kg. In other aspects, an IL-7 protein is administered at a dose of between about 700 pg/kg and about 800 pg/kg.
  • an IL-7 protein is administered at a dose of between about 800 pg/kg and about 900 pg/kg. In some aspects, an IL-7 protein is administered at a dose of between about 750 pg/kg and about 850 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of between about 850 pg/kg and about 950 pg/kg.
  • an IL-7 protein is administered at a dose of about 650 pg/kg, about 680 pg/kg, about 700 pg/kg, about 720 pg/kg, about 740 pg/kg, about 750 pg/kg, about 760 pg/kg, about 780 pg/kg, about 800 pg/kg, about 820 pg/kg, about 840 pg/kg, about 850 pg/kg, about 860 pg/kg, about 880 pg/kg, about 900 pg/kg, about 920 pg/kg, about 940 pg/kg, about 950 pg/kg, about 960 pg/kg, about 980 pg/kg, about 1,000 pg/kg, about 1,020 pg/kg, about 1,020 pg/kg, about 1,040 pg/kg, about 1,060 pg/kg, about 1,080 pg/kg
  • an IL-7 protein is administered at a dose of about 650 pg/kg. In other aspects, an IL-7 protein disclosed herein is administered at a dose of about 680 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 700 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 720 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 740 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 750 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 760 pg/kg.
  • an IL-7 protein is administered at a dose of about 780 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 800 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 820 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 840 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 850 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 860 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 880 pg/kg.
  • an IL-7 protein is administered at a dose of about 900 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 920 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 940 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 950 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 960 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 980 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,000 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,020 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,040 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,060 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,080 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,100 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,120 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,140 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,160 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,180 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,200 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,220 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,240 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,260 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,280 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,300 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,320 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,340 pg/kg. In some aspects, an IL- 7 protein is administered at a dose of about 1,360 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,380 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,400 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,420 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,440 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,460 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,480 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,500 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,520 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,540 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,560 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,580 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,600 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,620 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,640 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,660 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,680 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,700 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,720 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,740 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,760 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,780 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,800 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,820 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,840 pg/kg.
  • an IL-7 protein is administered at a dose of about 1,860 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,880 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,900 pg/kg. In certain aspects, an IL-7 protein is administered at a dose of about 1,920 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 1,940 pg/kg. In some aspects, an IL-7 protein is administered at a dose of about 1,960 pg/kg. In other aspects, an IL-7 protein is administered at a dose of about 1,980 pg/kg. In further aspects, an IL-7 protein is administered at a dose of about 2,000 pg/kg.
  • an IL-7 protein is administered at a dosing frequency of about once a week, about once in two weeks, about once in three weeks, about once in four weeks, about once in five weeks, about once in six weeks, about once in seven weeks, about once in eight weeks, about once in nine weeks, about once in 10 weeks, about once in 11 weeks, or about once in 12 weeks.
  • an IL-7 protein is administered at a dosing frequency of about once every 10 days, about once every 20 days, about once every 30 days, about once every 40 days, about once every 50 days, about once every 60 days, about once every 70 days, about once every 80 days, about once every 90 days, or about once every 100 days.
  • the IL-7 protein is administered once in three weeks.
  • the IL-7 protein is administered once a week. In some aspects, the IL-7 protein is administered once in two weeks. In certain aspects, the IL-7 protein is administered once in three weeks. In some aspects, the IL-7 protein is administered once in four weeks. In certain aspects, the IL-7 protein is administered once in six weeks. In further aspects, the IL-7 protein is administered once in eight weeks. In some aspects, the IL-7 protein is administered once in nine weeks. In certain aspects, the IL-7 protein is administered once in 12 weeks. In some aspects, the IL-7 protein is administered once every 10 days. In certain aspects, the IL-7 protein is administered once every 20 days. In other aspects, the IL-7 protein is administered once every 30 days. In some aspects, the IL-7 protein is administered once every 40 days.
  • the IL-7 protein is administered once every 50 days. In some aspects, the IL-7 protein is administered once every 60 days. In further aspects, the IL-7 protein is administered once every 70 days. In some aspects, the IL-7 protein is administered once every 80 days. In certain aspects, the IL-7 protein is administered once every 90 days. In some aspects, the IL-7 protein is administered once every 100 days.
  • the IL-7 protein is administered twice or more times in an amount of about 720 pg/kg at an interval of about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some aspects, the IL-7 protein is administered twice or more times in an amount of about 840 pg/kg at an interval of about 2 weeks, about 3 weeks, about 4 weeks, or about 5 weeks. In some aspects, the IL-7 protein is administered twice or more times in an amount of about 960 pg/kg at an interval of about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, or about 6 weeks.
  • the IL-7 protein is administered twice or more times in an amount of about 1200 pg/kg at an interval of about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or about 8 weeks. In some aspects, the IL-7 protein is administered twice or more times in an amount of about 1440 pg/kg at an interval of about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, about 8 weeks, about 10 weeks, about 12 weeks, or about 3 months.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once a week.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once a week. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once a week.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once a week. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once a week. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once a week. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once a week.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once a week. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once a week.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in two weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in two weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in two weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in two weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in two weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in two weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in two weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in two weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in two weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in three weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in three weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in three weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in three weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in three weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in three weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in three weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in three weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in four weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in four weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in four weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in four weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in four weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in four weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in four weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in four weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in four weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in five weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in five weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in five weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in five weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in five weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in five weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in five weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in five weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in five weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in six weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in six weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in six weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in six weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in six weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in six weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in six weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in six weeks. In some aspects, the IL- 7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in six weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in seven weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in seven weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in seven weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in seven weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in seven weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in seven weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in seven weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in seven weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in seven weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in eight weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 1 ,400 pg/kg with a dosing frequency of once in eight weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in eight weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in eight weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in eight weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in eight weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in eight weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in eight weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in eight weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in nine weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in three weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in three weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in nine weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in three weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in three weeks. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in three weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in three weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in nine weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in nine weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in 10 weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in 10 weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in 10 weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in 10 weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in 10 weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL- 7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in 10 weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in 10 weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in 10 weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in 10 weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in 11 weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in 11 weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in 11 weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in 11 weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in 11 weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL- 7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in 11 weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in 11 weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in 11 weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in 11 weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once in 12 weeks.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once in 12 weeks. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once in 12 weeks.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once in 12 weeks. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once in 12 weeks. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL- 7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once in 12 weeks. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once in 12 weeks.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once in 12 weeks. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once in 12 weeks.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 10 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 10 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 10 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 10 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 10 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 10 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 10 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 10 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 10 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 20 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 20 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 20 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 20 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 20 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 20 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 20 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 20 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 20 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 30 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 30 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 30 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 30 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 30 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 30 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 30 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 30 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 30 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 40 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 40 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 40 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 40 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 40 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 40 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 40 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 40 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 40 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 50 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 50 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 50 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 50 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 50 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 50 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 50 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 50 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 50 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 60 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 60 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 60 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 60 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 60 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 60 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 60 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 60 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 60 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 70 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 70 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 70 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 70 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 70 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 70 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 70 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 70 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 70 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 80 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 80 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 80 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 80 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 80 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 80 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 80 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 80 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 80 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 90 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 90 days. In other aspects, the IL-7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 90 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 90 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 90 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 90 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 90 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 90 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 90 days.
  • the IL-7 protein is administered at a dose of 60 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 120 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 240 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 480 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 720 pg/kg with a dosing frequency of once every 100 days.
  • the IL-7 protein is administered at a dose of 960 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 1,200 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 1,300 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 1,400 pg/kg with a dosing frequency of once every 100 days. In other aspects, the IL- 7 protein is administered at a dose of 1,420 pg/kg with a dosing frequency of once every 100 days.
  • the IL-7 protein is administered at a dose of 1,440 pg/kg with a dosing frequency of once every 100 days. In further aspects, the IL-7 protein is administered at a dose of 1,460 pg/kg with a dosing frequency of once every 100 days. In certain aspects, the IL-7 protein is administered at a dose of 1,480 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 1,500 pg/kg with a dosing frequency of once every 100 days. In further aspects, the IL-7 protein is administered at a dose of 1,600 pg/kg with a dosing frequency of once every 100 days.
  • the IL-7 protein is administered at a dose of 1,700 pg/kg with a dosing frequency of once every 100 days. In some aspects, the IL-7 protein is administered at a dose of 2,000 pg/kg with a dosing frequency of once every 100 days.
  • the nucleotide vaccine is a DNA vaccine.
  • the nucleotide vaccine is a mRNA vaccine.
  • IL-7 is administered to a subject (e.g., suffering from a tumor) as a protein (IL-7 protein), nucleic acid encoding the IL-7 protein, or both.
  • a subject e.g., suffering from a tumor
  • IL-7 protein a protein
  • nucleic acid encoding the IL-7 protein or both.
  • a nucleotide vaccine e.g., encoding a tumor antigen
  • IL-7 described herein
  • the nucleotide vaccine and/or IL-7 is administered to the subject parenthetically, intramuscularly, cutaneously, subcutaneously, ophthalmic, intravenously, intraperitoneally, intradermally, intraorbitally, intracerebrally, intracranially, intraspinally, intraventricular, intrathecally, intraci stemally, intracapsularly, or intratumorally.
  • methods disclosed herein can be used in combination with one or more additional therapeutic agent (e.g., anti-cancer and/or immunomodulating agents).
  • additional therapeutic agent e.g., anti-cancer and/or immunomodulating agents.
  • agents can include, for example, chemotherapy drugs, small molecule drugs, or antibodies that stimulate the immune response to a given cancer.
  • the methods described herein are used in combination with a standard of care treatment (e.g, surgery, radiation, and chemotherapy).
  • the one or more additional therapeutic agent comprises an immuno-oncology agent, such that multiple elements of the immune pathway can be targeted.
  • Non-limiting of such combinations include: a therapy that enhances tumor antigen presentation (e.g., dendritic cell vaccine, GM-CSF secreting cellular vaccines, CpG oligonucleotides, imiquimod); a therapy that inhibits negative immune regulation e.g., by inhibiting CTLA-4 and/or PD-1/PD-L1/PD-L2 pathway and/or depleting or blocking Tregs or other immune suppressing cells (e.g., myeloid-derived suppressor cells); a therapy that stimulates positive immune regulation, e.g., with agonists that stimulate the CD-137, OX-40, and/or CD40 or GITR pathway and/or stimulate T cell effector function; a therapy that increases systemically the frequency of anti-tumor T cells; a therapy that depletes or inhibits Tregs, such as Tregs in the tumor, e.g., using an antagonist of CD25 (e.g., daclizumab) or by ex vivo anti-CD25
  • immune checkpoint inhibitors that can be used in the present methods comprise a CTLA-4 antagonist (e.g., anti- CTLA-4 antibody), PD-1 antagonist (e.g., anti-PD-1 antibody, anti-PD-Ll antibody), TIM-3 antagonist (e.g., anti-TIM-3 antibody), or combinations thereof.
  • an immuno-oncology agent comprises an immune checkpoint activator (z.e., promotes signaling through the particular immune checkpoint pathway).
  • immune checkpoint activator comprises 0X40 agonist (e.g., anti-OX40 antibody), LAG-3 agonist (e.g. anti-LAG-3 antibody), 4-1BB (CD137) agonist (e.g., anti- CD137 antibody), GITR agonist (e.g., anti-GITR antibody), or any combination thereof.
  • IL-7 Proteins Useful for the Disclosure [0165] Disclosed herein are IL-7 proteins that can be used in combination with a nucleotide vaccine (e.g., encoding a tumor antigen), e.g., to treat a tumor (or cancer).
  • a nucleotide vaccine e.g., encoding a tumor antigen
  • IL- 7 protein useful for the present uses can be wild-type IL-7 or modified IL-7 (/. ⁇ ., not wild-type IL-7 protein) (e.g., IL-7 variant, IL-7 functional fragment, IL-7 derivative, or any combination thereof, e.g, fusion protein, chimeric protein, etc.) as long as the IL-7 protein contains one or more biological activities of IL-7, e.g., capable of binding to IL-7R, e.g., inducing early T-cell development, promoting T-cell homeostasis. See ElKassar and Gress. J Immunotoxicol . 2010 Mar; 7(1): 1-7.
  • an IL-7 protein of the present disclosure is not a wild-type IL- 7 protein (i.e., comprises one or more modifications).
  • modifications can include an oligopeptide and/or a half-life extending moiety. See WO 2016/200219, which is herein incorporated by reference in its entirety.
  • IL-7 binds to its receptor which is composed of the two chains IL-7Ra (CD127), shared with the thymic stromal lymphopoietin (TSLP) (Ziegler and Liu, 2006), and the common y chain (CD 132) for IL-2, IL- 15, IL-9 and IL-21. Whereas yc is expressed by most hematopoietic cells, IL-7Ra is nearly exclusively expressed on lymphoid cells. After binding to its receptor, IL-7 signals through two different pathways: Jak-Stat (Janus kinase-Signal transducer and activator of transcription) and PI3K/Akt responsible for differentiation and survival, respectively.
  • Jak-Stat Jak-Stat
  • PI3K/Akt PI3K/Akt responsible for differentiation and survival, respectively.
  • mice lack T-, B-, and NK-T cells.
  • IL-7a-/- mice (Peschon et al., 1994) have a similar but more severe phenotype than IL-7-/- mice (von Freeden- Jeffry et al., 1995), possibly because TSLP signaling is also abrogated in IL-7a-/- mice.
  • IL-7 is required for the development of y6 cells (Maki et al., 1996) and NK-T cells (Boesteanu et al., 1997).
  • an IL-7 protein useful for the present disclosure comprises an amino acid sequence as set forth in any one of SEQ ID NOs: 1 to 6.
  • the IL-7 protein comprises an amino acid sequence having a sequence identity of about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% or higher, to a sequence of SEQ ID NOS: 1 to 6.
  • the IL-7 protein includes a modified IL-7 or a fragment thereof, wherein the modified IL-7 or the fragment retains one or more biological activities of wildtype IL-7.
  • the IL-7 protein can be derived from humans, rats, mice, monkeys, cows, or sheep.
  • the human IL-7 can have an amino acid sequence represented by SEQ ID NO: 1 (Genbank AccessionNo. P13232); the rat IL-7 can have an amino acid sequence represented by SEQ ID NO: 2 (Genbank Accession No. P56478); the mouse IL-7 can have an amino acid sequence represented by SEQ ID NO: 3 (Genbank Accession No. P10168); the monkey IL-7 can have an amino acid sequence represented by SEQ ID NO: 4 (Genbank Accession No. NP 001279008); the cow IL-7 can have an amino acid sequence represented by SEQ ID NO: 5 (Genbank Accession No. P26895), and the sheep IL-7 can have an amino acid sequence represented by SEQ ID NO: 6 (Genbank Accession No. Q28540).
  • an IL-7 protein useful for the present disclosure comprises an IL- 7 fusion protein.
  • an IL-7 fusion protein comprises (i) an oligopeptide and (i) an IL-7 or a variant thereof.
  • the oligopeptide is linked to the N-terminal region of the IL-7 or a variant thereof.
  • an oligopeptide disclosed herein consists of 1 to 10 amino acids. In certain aspects, an oligopeptide consists of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or 10 amino acids. In some aspects, one or more amino acids of an oligopeptide are selected from the group consisting of methionine, glycine, and combinations thereof.
  • an oligopeptide is selected from the group consisting of methionine (M), glycine (G), methionine-methionine (MM), glycine-glycine (GG), methionine-glycine (MG), glycine-methionine (GM), methionine-methionine- methionine (MMM), methionine-methionine-glycine (MMG), methionine-glycine-methionine (MGM), glycine-methionine-methionine (GMM), methionine-glycine-glycine (MGG), glycine-methionine-glycine (GMG), glycine-methionine-glycine (GGM), glycine-glycine-methionine (GGM), glycine-glycine-methionine (GGM), glycine-glycine-methionine (GGM), glycine-glycine-
  • an IL-7 fusion protein comprises (i) an IL-7 or a variant thereof, and (ii) a half-life extending moiety.
  • a half-life extending moiety extends the half-life of the IL-7 or variant thereof.
  • a half-life extending moiety is linked to the C-terminal region of an IL-7 or a variant thereof.
  • an IL-7 fusion protein comprises (i) IL-7 (a first domain), (ii) a second domain that includes an amino acid sequence having 1 to 10 amino acid residues consisting of methionine, glycine, or a combination thereof, e.g., MGM, and (iii) a third domain comprising a half-life extending moiety.
  • the half-life extending moiety can be linked to the N-terminal or the C-terminal of the first domain or the second domain.
  • the IL-7 including the first domain and the second domain can be linked to both terminals of the third domain.
  • Non-limiting examples of half-life extending moieties include: Fc, albumin, an albumin-binding polypeptide, Pro/Ala/Ser (PAS), a C-terminal peptide (CTP) of the P subunit of human chorionic gonadotropin, polyethylene glycol (PEG), long unstructured hydrophilic sequences of amino acids (XTEN), hydroxyethyl starch (HES), an albumin-binding small molecule, and combinations thereof.
  • a half-life extending moiety is Fc.
  • Fc is from a modified immunoglobulin in which the antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) weakened due to the modification in the binding affinity with the Fc receptor and/or a complement.
  • the modified immunoglobulin can be selected from the group consisting of IgGl, IgG2, IgG3, IgG4, IgAl, IgA2, IgD, IgE, and a combination thereof.
  • an Fc is a hybrid Fc ("hFc" or "hyFc"), comprising a hinge region, a CH2 domain, and a CH3 domain.
  • a hinge region of a hybrid Fc disclosed herein comprises a human IgD hinge region.
  • a CH2 domain of a hybrid Fc comprises a part of human IgD CH2 domain and a part of human IgG4 CH2 domain.
  • a CH3 domain of a hybrid Fc comprises a part of human IgG4 CH3 domain.
  • a hybrid Fc disclosed herein comprises a hinge region, a CH2 domain, and a CH3 domain, wherein the hinge region comprises a human IgD hinge region, wherein the CH2 domain comprises a part of human IgD CH2 domain and a part of human IgG4 CH2 domain, and wherein the CH3 domain comprises a part of human IgG4 CH3 domain.
  • an Fc disclosed herein can be an Fc variant.
  • the term "Fc variant” refers to an Fc which was prepared by substituting a part of the amino acids among the Fc region or by combining the Fc regions of different kinds.
  • the Fc region variant can prevent from being cut off at the hinge region.
  • a Fc variant comprises modifications at the 144 th amino acid and/or 145 th amino acid of SEQ ID NO: 9.
  • the 144 th amino acid (K) and/or the 145 th amino acid (K) is substituted with G or S.
  • an Fc or an Fc variant disclosed herein can be represented by the following formula: N' - (Zl)p - Y - Z2 - Z3 - Z4 - C, wherein:
  • N' comprises the N-terminus
  • Z1 comprises an amino acid sequence having 5 to 9 consecutive amino acid residues from the amino acid residue at position 98 toward the N-terminal, among the amino acid residues at positions from 90 to 98 of SEQ ID NO: 7;
  • Y comprises an amino acid sequence having 5 to 64 consecutive amino acid residues from the amino acid residue at position 162 toward the N-terminal, among the amino acid residues at positions from 99 to 162 of SEQ ID NO: 7;
  • Z2 comprises an amino acid sequence having 4 to 37 consecutive amino acid residues from the amino acid residue at position 163 toward the C-terminal, among the amino acid residues at positions from 163 to 199 of SEQ ID NO: 7;
  • Z3 comprises an amino acid sequence having 71 to 106 consecutive amino acid residues from the amino acid residue at position 220 toward the N-terminal, among the amino acid residues at positions from 115 to 220 of SEQ ID NO: 8;
  • Z4 comprises an amino acid sequence having 80 to 107 consecutive amino acid residues from the amino acid residue at position 221 toward the C-terminal, among the amino acid residues at positions from 221 to 327 of SEQ ID NO: 8.
  • a Fc region disclosed herein can include the amino acid sequence of SEQ ID NO: 9 (hyFc), SEQ ID NO: 10 (hyFcMl), SEQ ID NO: 11 (hyFcM2), SEQ ID NO: 12 (hyFcM3), or SEQ ID NO: 13 (hyFcM4).
  • the Fc region can include the amino acid sequence of SEQ ID NO: 14 (a non-lytic mouse Fc).
  • Fc regions that can be used with the present disclosure are described in U.S. Pat. No. 7,867,491, which is herein incorporated by reference in its entirety.
  • an IL-7 fusion protein disclosed herein comprises both an oligopeptide and a half-life extending moiety.
  • an IL-7 protein can be fused to albumin, a variant, or a fragment thereof.
  • examples of the IL-7-albumin fusion protein can be found at International Application Publication No. WO 2011/124718 AL
  • an IL-7 protein is fused to a pre-pro-B cell Growth Stimulating Factor (PPBSF), optionally by a flexible linker.
  • PBSF pre-pro-B cell Growth Stimulating Factor
  • an IL-7 protein useful for the disclosure is an IL-7 conformer that has a particular three dimensional structure.
  • an IL-7 protein can be fused to an Ig chain, wherein amino acid residues 70 and 91 in the IL-7 protein are glycosylated the amino acid residue 116 in the IL-7 protein is non- glycosylated. See US 7,323,549 B2.
  • an IL-7 protein that does not contain potential T-cell epitopes (thereby to reduce anti-IL-7 T-cell responses) can also be used for the present disclosure. See US 2006/0141581 AL
  • an IL-7 protein that has one or more amino acid residue mutations in carboxy-terminal helix D region can be used for the present disclosure.
  • the IL-7 mutant can act as IL-7R partial agonist despite lower binding affinity for the receptor. See US 2005/0054054A1. Any IL-7 proteins described in the above listed patents or publications are incorporated herein by reference in their entireties.
  • IL-7 proteins useful for the present disclosure are described in US 7708985, US 8034327, US 8153114, US 7589179, US 7323549, US 7960514, US 8338575, US 7118754, US 7488482, US 7670607, US 6730512, W00017362, GB2434578A, WO 2010/020766 A2, WO91/01143, Beq et al., Blood, vol. 114 (4), 816, 23 July 2009, Kang et al., J. Virol. Doi: 10.1128/JVI.02768-15, Martin et al., Blood, vol.
  • an oligopeptide disclosed herein is directly linked to the N-terminal region of IL-7 or a variant thereof. In some aspects, an oligopeptide is linked to the N-terminal region via a linker. In some aspects, a half-life extending moiety disclosed herein is directly linked to the C-terminal region of IL-7 or a variant thereof. In certain aspects, a half-life extending moiety is linked to the C-terminal region via a linker. In some aspects, a linker is a peptide linker. In certain aspects, a peptide linker comprises a peptide of 10 to 20 amino acid residues consisting of Gly and Ser residues.
  • a linker is an albumin linker.
  • a linker is a chemical bond.
  • a chemical bond comprises a disulfide bond, a diamine bond, a sulfide-amine bond, a carboxy-amine bond, an ester bond, a covalent bond, or combinations thereof.
  • the linker is a peptide linker, in some aspects, the connection can occur in any linking region. They can be coupled using a crosslinking agent known in the art.
  • examples of the crosslinking agent can include N-hydroxy succinimide esters such as l,l-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, and 4- azidosalicylic acid; imido esters including disuccinimidyl esters such as 3,3'-dithiobis (succinimidyl propionate), and bifunctional maleimides such as bis-Nmaleimido-l,8-octane, but is not limited thereto.
  • succinimide esters such as l,l-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, and 4- azidosalicylic acid
  • imido esters including disuccinimidyl esters such as 3,3'-dithiobis (succinimidyl propionate), and bifunctional maleimides such as bis-Nmaleimido-l,8-octane, but is not limited
  • an IL-7 (or variant thereof) portion of IL-7 fusion protein disclosed herein comprises an amino sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 98%, or at least 99% identical to an amino acid sequence set forth in SEQ ID NOs: 15-20.
  • an IL-7 (or variant thereof) portion of IL-7 fusion protein disclosed herein comprises the amino acid sequence set forth in SEQ ID NOs: 15-20.
  • an IL-7 fusion protein comprises: a first domain including a polypeptide having the activity of IL-7 or a similar activity thereof; a second domain comprising an amino acid sequence having 1 to 10 amino acid residues consisting of methionine, glycine, or a combination thereof; and a third domain, which is an Fc region of modified immunoglobulin, coupled to the C-terminal of the first domain.
  • an IL-7 fusion protein of the present disclosure comprises the amino acid sequence set forth in SEQ ID NOs: 21-25.
  • an IL- 7 fusion protein disclosed herein comprises the amino acid sequence set forth in SEQ ID NOs: 26 and 27.
  • an IL-7 protein useful for the present disclosure can increase absolute lymphocyte counts in a subject when administered to the subject.
  • the subject suffers from a disease or disorder described herein (e.g., cancer).
  • the subject is a healthy individual (e.g., does not suffer from a disease or disorder described herein, e.g., cancer).
  • the absolute lymphocyte count is increased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% or more, compared to a reference (e.g., corresponding level in a subject that did not receive the IL-7 protein).
  • a reference e.g., corresponding level in a subject that did not receive the IL-7 protein
  • nucleotide vaccines can comprise one or more vectors that include one or more heterologous nucleic acids encoding a tumor antigen.
  • nucleotide vaccines can further comprise one or more vectors that include one or more heterologous nucleic acids encoding an additional agent, e.g., IL-7 protein disclosed herein.
  • the tumor antigen and the IL-7 protein can be encoded in a single vector. In some aspects, the tumor antigen and the IL-7 protein are encoded in separate vectors.
  • nucleotide vaccines disclosed herein can encode any antigen or protein known in the art that can be useful in treating a tumor (or cancer). As described herein, in some aspects, the nucleotide vaccine encodes a tumor antigen.
  • tumor antigens include Lrrc27, Plekhol, Pttgl, Xpo4, Exoc4, Pank3, TmemlOl, Map3k6, Met, BC057079, Histlh3e, Prkagl, Neil3, guanylate cyclase C (GC-C), epidermal growth factor receptor (EGFR or erbB-1), human epidermal growth factor receptor 2 (HER2 or erbB2), erbB-3, erbB-4, MUC-1, melanoma-associated chondroitin sulfate proteoglycan (MCSP), mesothelin (MSLN), folate receptor 1 (FOLR1), CD4, CD19, CD20, CD22, CD30, CD33, CD38, CD44, CD44v6, CD44v7/8, CD70, CD 123, CD 138, CD171, CEA, CSPG4, CXCR5, c-Met, HERV-envelope protein, erios
  • the tumor antigen comprises a cancer neoantigen (/. ⁇ ., mutated antigens specifically expressed by tumor tissue and not expressed on the surface of normal cells).
  • cancer neoantigen /. ⁇ ., mutated antigens specifically expressed by tumor tissue and not expressed on the surface of normal cells.
  • antigens including methods of identification, are known in the art. See, e.g., Hutchison et al.,Mamm Genome 29(11): 714-730 (Aug. 2018), which is incorporated herein by reference in its entirety.
  • nucleotides vaccines of the present disclosure encodes at least about one, at least about two, at least about three, at least about four, at least about five, at least about six, at least about seven, at least about eight, at least about nine, at least about ten, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, at least bout 27, at least about 28, at least about 29, at least about 30, at least about 35, at least about 40, at least about 45, or at least about 50 or more different tumor antigens.
  • nucleotide vaccines of the present disclosure comprise both DNA vaccine and mRNA vaccine.
  • DNA vaccines, including methods of making are described, for example, in U.S. Pat. Nos. 7,795,017 B2 and 5,643,578 A, each of which is herein incorporated by reference in its entirety.
  • mRNA vaccines, including methods of making are described, for example in U.S. Publication Nos. 2018/0311336 Al and 2020/0085852 Al, each of which is herein incorporated by reference in its entirety.
  • CMV cytomegalovirus
  • Another factor known to affect the immune response elicited by nucleotide vaccine immunization is the method of delivery. For instance, parenteral routes can yield low rates of gene transfer and produce considerable variability of gene expression (Montgomery et al., DNA Cell Bio., 12:777-783 (1993)). High- velocity inoculation of plasmids (e.g., DNA plasmids), using a gene-gun, have been shown to enhance immune responses in mice (Fynan et al., Proc.
  • Vectors containing the nucleotide vaccines of the present disclosure can also be introduced into the desired host by other methods known in the art, e.g., transfection, electroporation, microinjection, transduction, cell fusion, DEAE dextran, calcium phosphate precipitation, lipofection (lysosome fusion), or a DNA vector transporter (see, e.g., Wu etal.,J Biol. Chem.
  • the one or more vectors used in constructing the nucleotide vaccines of the present disclosure comprises expression vectors, viral vectors, plasmid vectors, or combinations thereof.
  • the vector is an expression vector.
  • an "expression vector” refers to any nucleic acid construct which contains the necessary elements for the transcription and translation of an inserted coding sequence, or in the case of an RNA viral vector, the necessary elements for replication and translation, when introduced into an appropriate host cell.
  • Expression vectors can include plasmids, phagemids, viruses, and derivatives thereof. Once the expression vector is inside the cell, the protein that is encoded by the gene can be produced by the cellular-transcription and translation machinery ribosomal complexes.
  • the plasmid can be engineered to contain regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene carried on the expression vector.
  • the nucleotide vaccines described herein can comprise a circular plasmid or a linear nucleic acid.
  • the circular plasmid and linear nucleic acid are capable of directing expression of a particular heterologous nucleotide sequence (e.g., encoding a tumor antigen and/or IL-7 protein described herein) in an appropriate subject cell.
  • the vector can have a promoter operably linked to the nucleotide sequence (e.g., encoding a tumor antigen and/or IL-7 protein described herein), which can be operably linked to termination signals.
  • the vector can also contain sequences required for proper translation of the nucleotide sequence (e.g., encoding a tumor antigen and/or IL-7 protein described herein).
  • the vector comprising the nucleotide sequence of interest e.g., encoding a tumor antigen and/or IL-7 protein described herein
  • the expression of the nucleotide sequence (e.g., encoding a tumor antigen and/or IL-7 protein described herein) in the expression cassette can be under the control of a constitutive promoter or an inducible promoter, which initiates transcription only when the host cell is exposed to some particular external stimulus.
  • the promoter can also be specific to a particular tissue or organ or stage of development.
  • the nucleotide vaccine comprises a circular plasmid, which can transform a target cell by integration into the cellular genome or exist extrachromosomally (e.g., autonomous replicating plasmid with an origin of replication).
  • the vector can be pVAX, pcDNA3.0, provax, or any other expression vector capable of expressing a heterologous nucleic acid (e.g, encoding a tumor antigen and/or IL-7 protein described herein) and enabling a cell to translate the sequence such that is recognized by the immune system.
  • the nucleotide vaccine comprises a linear nucleic acid (e.g, encoding a tumor antigen and/or IL-7 protein described herein) that is capable of being efficiently delivered to a subject and expressing one or more desired proteins (e.g., tumor antigen and/or IL-7 protein disclosed herein).
  • the linear nucleic acid can contain a promoter, an intron, a stop codon, and/or a polyadenylation signal, which help regulate the expression of the desired proteins (e.g., tumor antigen and/or IL-7 protein disclosed herein).
  • the linear nucleic acid does not contain any antibiotic resistance genes and/or a phosphate backbone.
  • the linear nucleic acid does not contain other nucleic acid sequences unrelated to the desired protein (e.g., tumor antigen and/or IL-7 protein disclosed herein) expression.
  • the linear nucleic acid e.g., tumor antigen and/or IL-7 protein disclosed herein
  • the linear nucleic acid can be derived from any plasmid capable of being linearized.
  • Non-limiting examples of plasmids that can be used include pNP (Puerto Rico/34), pM2 (New Caledonia/99), WLV009, pVAX, pcDNA3.0, provax, or combinations thereof.
  • vectors useful for constructing the nucleotide vaccines of the present disclosure can comprise a promoter.
  • the promoter can be any promoter that is capable of driving gene expression and regulating expression of the nucleic acid (e.g., encoding a tumor antigen and/or IL-7 protein disclosed herein).
  • the promoter is a cis-acting sequence element required for transcription via a DNA dependent RNA polymerase. Selection of the promoter used to direct expression of a heterologous nucleic acid (e.g., encoding a tumor antigen and/or IL-7 protein disclosed herein) can depend on the particular application.
  • the promoter can be a CMV promoter, SV40 early promoter, SV40 later promoter, metallothionein promoter, murine mammary tumor virus promoter, Rous sarcoma virus promoter, polyhedrin promoter, or another promoter shown effective for expression in eukaryotic cells.
  • CMV promoter SV40 early promoter
  • SV40 later promoter metallothionein promoter
  • murine mammary tumor virus promoter murine mammary tumor virus promoter
  • Rous sarcoma virus promoter Rous sarcoma virus promoter
  • polyhedrin promoter or another promoter shown effective for expression in eukaryotic cells.
  • promoters that are useful for the present disclosure are described in U.S. Pat. No. 7,557,200 B2, which is herein incorporated by reference in its entirety.
  • the vectors that can be used with the present disclosure include an enhancer and an intron with functional splice donor and acceptor sites.
  • the vector can contain a transcription termination region downstream of the structural gene to provide for efficient termination.
  • the termination region can be obtained from the same gene as the promoter sequence or can be obtained from different genes.
  • the vector is a viral vector.
  • viral vectors include, but are not limited to, nucleic acid sequences from the following viruses: retrovirus, such as Moloney murine leukemia virus, Harvey murine sarcoma virus, murine mammary tumor virus, and Rous sarcoma virus; lentivirus; adenovirus; adeno-associated virus; SV40-type viruses; polyomaviruses; Epstein-Barr viruses; papilloma viruses; herpes virus; vaccinia virus; polio virus; and RNA virus such as a retrovirus.
  • retrovirus such as Moloney murine leukemia virus, Harvey murine sarcoma virus, murine mammary tumor virus, and Rous sarcoma virus
  • retrovirus such as Moloney murine leukemia virus, Harvey murine sarcoma virus, murine mammary tumor virus, and Rous sarcoma virus
  • lentivirus such as Moloney murine leukemia virus
  • Non-cytopathic viral vectors are based on non-cytopathic eukaryotic viruses in which non- essential genes have been replaced with the gene of interest.
  • Non-cytopathic viruses include retroviruses, the life cycle of which involves reverse transcription of genomic viral RNA into DNA with subsequent proviral integration into host cellular DNA.
  • a vector is derived from an adeno-associated virus.
  • a vector is derived from a lentivirus. Examples of the lentiviral vectors are disclosed in WO9931251, W09712622, W09817815, W09817816, and WO9818934, each which is incorporated herein by reference in its entirety.
  • a method for making a therapeutic agent disclosed herein e.g., an IL-7 protein
  • a method for making a therapeutic agent disclosed herein can comprise expressing the therapeutic agent (e.g., an IL-7 protein) in a cell comprising a nucleic acid molecule encoding the therapeutic agent, e.g., SEQ ID NOs: 29-39. Additional details regarding the method for making an IL-7 protein disclosed herein are provided, e.g., in U.S. Publ. No. 2018/0273596 Al, which is herein incorporated by reference in its entirety. Host cells comprising these nucleotide sequences are encompassed herein.
  • Non-limiting examples of host cell that can be used include immortal hybridoma cell, NS/0 myeloma cell, 293 cell, Chinese hamster ovary (CHO) cell, HeLa cell, human amniotic fluid-derived cell (CapT cell), COS cell, or combinations thereof.
  • immortal hybridoma cell NS/0 myeloma cell
  • 293 cell Chinese hamster ovary (CHO) cell
  • HeLa cell human amniotic fluid-derived cell
  • CapT cell human amniotic fluid-derived cell
  • COS cell or combinations thereof.
  • compositions comprising one or more therapeutic agents (e.g., a nucleotide vaccine and/or IL-7) having the desired degree of purity in a physiologically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA).
  • a composition disclosed herein comprises one or more nucleotide vaccines encoding a tumor antigen.
  • a composition disclosed herein comprises an IL-7 (e.g., those disclosed herein).
  • such compositions can be used in combination (e.g., a first composition comprising a nucleotide vaccine, and a second composition comprising an IL-7).
  • composition comprising an IL-7 is administered after administering the composition comprising the nucleotide vaccine (e.g., after the peak expansion phase of the tumor-specific T cell immune response).
  • a composition comprises both (i) a nucleotide vaccine encoding a tumor antigen, and (ii) an IL-7.
  • Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, hist
  • composition disclosed herein comprises one or more additional components selected from: a bulking agent, stabilizing agent, surfactant, buffering agent, or combinations thereof.
  • Buffering agents useful for the current disclosure can be a weak acid or base used to maintain the acidity (pH) of a solution near a chosen value after the addition of another acid or base.
  • Suitable buffering agents can maximize the stability of the pharmaceutical compositions by maintaining pH control of the composition.
  • Suitable buffering agents can also ensure physiological compatibility or optimize solubility. Rheology, viscosity and other properties can also dependent on the pH of the composition.
  • Common buffering agents include, but are not limited to, a Tris buffer, a Tris-Cl buffer, a histidine buffer, a TAE buffer, a HEPES buffer, a TBE buffer, a sodium phosphate buffer, a MES buffer, an ammonium sulfate buffer, a potassium phosphate buffer, a potassium thiocyanate buffer, a succinate buffer, a tartrate buffer, a DIPSO buffer, a HEPPSO buffer, a POPSO buffer, a PIPES buffer, a PBS buffer, a MOPS buffer, an acetate buffer, a phosphate buffer, a cacodylate buffer, a glycine buffer, a sulfate buffer, an imidazole buffer, a guanidine hydrochloride buffer, a phosphate-citrate buffer, a borate buffer, a mal onate buffer, a 3 -picoline buffer, a 2-picoline buffer, a 4-picoline
  • a composition disclosed herein further comprises a bulking agent.
  • Bulking agents can be added to a pharmaceutical product in order to add volume and mass to the product, thereby facilitating precise metering and handling thereof.
  • Bulking agents that can be used with the present disclosure include, but are not limited to, sodium chloride (NaCl), mannitol, glycine, alanine, or combinations thereof.
  • composition disclosed herein can also comprise a stabilizing agent.
  • stabilizing agents that can be used with the present disclosure include: sucrose, trehalose, raffinose, arginine, or combinations thereof.
  • a composition disclosed herein comprises a surfactant.
  • the surfactant can be selected from the following: alkyl ethoxylate, nonylphenol ethoxylate, amine ethoxylate, polyethylene oxide, polypropylene oxide, fatty alcohols such as cetyl alcohol or oleyl alcohol, cocamide MEA, cocamide DEA, polysorbates, dodecyl dimethylamine oxide, or combinations thereof.
  • the surfactant is polysorbate 20 or polysorbate 80.
  • a composition comprising a nucleotide vaccine can be formulated using the same formulation used for formulating a composition comprising an IL-7.
  • a nucleotide vaccine and IL-7 are formulated using different formulations.
  • an IL-7 disclosed herein is formulated in a composition comprising (a) a basal buffer, (b) a sugar, and (c) a surfactant.
  • the basal buffer comprises histidine-acetate or sodium citrate.
  • the basal buffer is at a concentration of about 10 to about 50 nM.
  • a sugar comprises sucrose, trehalose, dextrose, or combinations thereof.
  • the sugar is present at a concentration of about 2.5 to about 5.0 w/v%.
  • the surfactant is selected from polysorbate, polyoxyethylene alkyl ether, polyoxyethylene stearate, alkyl sulfates, polyvinyl pyridone, poloxamer, or combinations thereof. In some aspects, the surfactant is at a concentration of about 0.05% to about 6.0 w/v%.
  • a composition disclosed herein (e.g., comprising a nucleotide vaccine and/or IL-7) further comprises an amino acid.
  • the amino acid is selected from arginine, glutamate, glycine, histidine, or combinations thereof.
  • the composition further comprises a sugar alcohol.
  • sugar alcohol includes: sorbitol, xylitol, maltitol, mannitol, or combinations thereof.
  • an IL-7 disclosed herein is formulated in a composition comprising the following: (a) sodium citrate (e.g., about 20 mM), (b) sucrose (e.g., about 5%), (c) sorbitol (e.g., about 1.5%), and (d) Tween 80 (e.g., about 0.05%).
  • an IL-7 is formulated as described in U.S. Publ. No. 2018/0327472 Al, which is incorporated herein in its entirety.
  • a pharmaceutical composition disclosed herein can be formulated for any route of administration to a subject.
  • routes of administration include intramuscularly, cutaneously, subcutaneously, ophthalmic, intravenously, intraperitoneally, intradermally, intraorbitally, intracerebrally, intracranially, intraspinally, intraventricular, intrathecally, intraci stemally, intracapsularly, or intratum orally.
  • Parenteral administration characterized by, e.g, cutaneous, subcutaneous, intramuscular, or intravenous injection, is also contemplated herein.
  • a nucleotide vaccine and IL-7 are administered using the same route of administration.
  • a nucleotide vaccine and IL-7 are administered using different routes of administration.
  • injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions.
  • the injectables, solutions and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol or ethanol.
  • compositions to be administered can also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrins.
  • auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrins.
  • Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents and other pharmaceutically acceptable substances.
  • aqueous vehicles include Sodium Chloride Injection, Ringers Injection, Isotonic Dextrose Injection, Sterile Water Injection, Dextrose and Lactated Ringers Injection.
  • Nonaqueous parenteral vehicles include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil and peanut oil.
  • Antimicrobial agents in bacteriostatic or fungistatic concentrations can be added to parenteral preparations packaged in multiple-dose containers which include phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride and benzethonium chloride.
  • Isotonic agents include sodium chloride and dextrose.
  • Buffers include phosphate and citrate.
  • Antioxidants include sodium bisulfate.
  • Local anesthetics include procaine hydrochloride.
  • Suspending and dispersing agents include sodium carboxymethylcelluose, hydroxypropyl methylcellulose and polyvinylpyrrolidone.
  • Emulsifying agents include Polysorbate 80 (TWEEN® 80).
  • a sequestering or chelating agent of metal ions includes EDTA.
  • Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol and propylene glycol for water miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.
  • Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders, ready to be combined with a solvent just prior to use, including hypodermic tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle just prior to use and sterile emulsions.
  • the solutions can be either aqueous or nonaqueous.
  • suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol and mixtures thereof.
  • PBS physiological saline or phosphate buffered saline
  • Topical mixtures comprising an antibody are prepared as described for the local and systemic administration.
  • the resulting mixture can be a solution, suspension, emulsions or the like and can be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, dermal patches or any other formulations suitable for topical administration.
  • a therapeutic agent described herein can be formulated as an aerosol for topical application, such as by inhalation (see, e.g., U.S. Patent Nos. 4,044,126, 4,414,209 and 4,364,923, which describe aerosols for delivery of a steroid useful for treatment of inflammatory diseases, particularly asthma).
  • These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a microfine powder for insufflations, alone or in combination with an inert carrier such as lactose.
  • the particles of the formulation can have diameters of less than about 50 microns, e.g., less than about 10 microns.
  • a therapeutic agent disclosed herein can be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intraci sternal or intraspinal application.
  • Topical administration is contemplated for transdermal delivery and also for administration to the eyes or mucosa, or for inhalation therapies.
  • Nasal solutions of the antibody alone or in combination with other pharmaceutically acceptable excipients can also be administered.
  • Transdermal patches including iontophoretic and electrophoretic devices, are well known to those of skill in the art, and can be used to administer a therapeutic agent (e.g., those disclosed herein).
  • a therapeutic agent e.g., those disclosed herein.
  • such patches are disclosed in U.S. Patent Nos. 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010715, 5,985,317, 5,983,134, 5,948,433, and 5,860,957, each of which is herein incorporated by reference in its entirety.
  • a pharmaceutical composition comprising a therapeutic agent described herein is a lyophilized powder, which can be reconstituted for administration as solutions, emulsions and other mixtures. It can also be reconstituted and formulated as solids or gels.
  • the lyophilized powder is prepared by dissolving an antibody or antigen-binding portion thereof described herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent.
  • the lyophilized powder is sterile.
  • the solvent can contain an excipient which improves the stability or other pharmacological component of the powder or reconstituted solution, prepared from the powder.
  • Excipients that can be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose or other suitable agent.
  • the solvent can also contain a buffer, such as citrate, sodium or potassium phosphate or other such buffer known to those of skill in the art at, in some aspects, about neutral pH.
  • sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides the desired formulation.
  • the resulting solution can be apportioned into vials for lyophilization. Each vial can contain a single dosage or multiple dosages of the compound.
  • the lyophilized powder can be stored under appropriate conditions, such as at about 4°C to room temperature.
  • Reconstitution of this lyophilized powder with water for injection provides a formulation for use in parenteral administration.
  • the lyophilized powder is added to sterile water or other suitable carrier. The precise amount depends upon the selected compound. Such amount can be empirically determined.
  • compositions provided herein can also be formulated to be targeted to a particular tissue, receptor, or other area of the body of the subject to be treated. Many such targeting methods are well known to those of skill in the art. All such targeting methods are contemplated herein for use in the instant compositions. For non-limiting examples of targeting methods, see, e.g., U.S. Patent Nos.
  • compositions to be used for in vivo administration can be sterile. This is readily accomplished by filtration through, e.g., sterile filtration membranes.
  • EXAMPLE 1 ANALYSIS OF ADMINISTRATION SCHEDULE OF IL-7 AND NUCLEOTIDE VACCINE COMBINATION THERAPY
  • DNA vaccines encoding one or more neoantigen epitopes from an estrogen-receptor positive murine breast cancer was constructed. Briefly, nucleic acids encoding one or more of the following epitopes were inserted into a single pcDNA3.1(+) backbone: Lrrc27, Plekhol, Pttgl, Xpo4, Exoc4, Pank3, TmemlOl, Map3k6, Met, BC057079, Histlh3e, Prkagl, and Neil3. See Hundal et al., Genome Med 8(1): 11 (Jan. 2016), which is herein incorporated by reference in its entirety.
  • naive C57BL6 mice were immunized with the DNA vaccine construct for a total of three doses (4 pg / dose) at a dosing frequency of once every three days. See FIG. 1 A.
  • the DNA vaccine was administered cutaneously to the mice using a gene gun. Some of the animals received a single dose of IL-7 (10 mg/kg) at either day 4 or day 13 post initial DNA vaccine administration.
  • the different treatment groups were as follows: (i) control vector only (Gl); (ii) DNA vaccine only (G2); (iii) DNA vaccine + IL-7 administration at day 4 post initial DNA vaccine administration (G3); and (iv) DNA vaccine + IL-7 administration at day 13 post initial DNA vaccine administration (G4).
  • Some of the animals from each of the treatment groups were sacrificed at days 11, 20, and 29 post initial DNA vaccine administration, and the tumor-specific T cell immune response was assessed in the spleen using an IFN-y ELISPOT assay.
  • the total number of tumor-specific T cells in the spleen of animals treated with IL-7 during the expansion phase was higher compared to animals that received the DNA vaccine alone (see FIG. ID).
  • day 20 post initial DNA vaccine administration there was no significant different in the tumor-specific T cell immune response (both in terms of frequency and total number) compared to animals treated with DNA vaccine alone (compare G2 and G3 in FIGs. IE and IF).
  • EXAMPLE 2 ANALYSIS OF THE DOSAGE EFFECT OF IL-7 ON THE EFFICACY OF IL-7 AND NUCLEOTIDE VACCINE COMBINATION THERAPY
  • IL-7 when administered in combination with a nucleotide vaccine disclosed herein, naive C57BL6 mice were immunized with the DNA vaccine construct as described in Example 1 (see FIG. 2A). Then, at day 13 post initial DNA vaccine administration (z.e., during the contraction phase), a single dose of IL-7 was administered to the animals at one of the following doses: 5, 10, or 15 mg/kg.
  • the different treatment groups were as follows: (i) control vector only (Gl); (ii) DNA vaccine only (G2); (iii) DNA vaccine + 5 mg/kg of IL-7 (G3); (iv) DNA vaccine + 10 mg/kg of IL-7 (G4); and (v) DNA vaccine + 15 mg/kg of IL-7 (G5).
  • Animals were sacrificed at day 20 post initial DNA vaccine administration, and the tumor-specific T cell immune response was assessed in the spleen and the lymph nodes using an IFN-y ELISPOT assay.
  • EXAMPLE 3 ANALYSIS OF THE ANTI-TUMOR EFFECTS OF A NUCLEOTIDE VACCINE AND IL-7 COMBINATION THERAPY [0238]
  • E0771 a syngenic breast cancer animal model
  • the animals were immunized with the DNA vaccine construct as described in Examples 1 and 2 (see FIG. 3A).
  • the animals were implanted subcutaneously with E0771 tumor cells (5 x 10 5 cells/mouse).
  • some of the animals received a single administration of IL-7 (5 mg/kg).
  • the treatment groups were as follows: (i) control vector alone (Gl); (ii) DNA vaccine alone (G2); and (iii) DNA vaccine + IL-7 (5 mg/kg) at day 13 post initial DNA vaccine administration. Then, tumor volume was measured in the animals periodically.
  • EXAMPLE 4 ANALYSIS OF THE EFFECT OF IL-7 ADMINISTRATION ON MEMORY T CELLS INDUCED AFTER NUCLEOTIDE VACCINE ADMINISTRATION
  • OVA ovalbumin
  • the animals will receive one of the following treatments: (i) no treatment; (ii) DNA-based OVA vaccine alone; (iii) DNA vaccine encoding a tumor antigen (e.g., such as those described in Example 1) alone; (iv) DNA vaccine encoding a tumor antigen + IL-7.
  • the IL-7 will be administered to the animals at a single dose of 5 mg/kg during the contraction phase of the tumor-specific T cell immune response (e.g., at day 13 post initial DNA vaccine administration).
  • mice At about day 60 post initial DNA vaccine administration, the mice will be sacrificed and mononuclear cells from the periphery (spleen, blood) and the bone marrow will be collected for immune monitoring.
  • EXAMPLE 5 ANALYSIS OF THE EFFECT OF IL-7 ADMINISTRATION ON T CELL- MEDIATED CYTOTOXICITY AFTER NUCLEOTIDE VACCINE ADMINISTRATION
  • mice were immunized with the DNA vaccine construct as described in Examples 1 and 2 (see FIG. 4A).
  • the DNA vaccine construct As described in Examples 1 and 2 (see FIG. 4A).
  • some of the animals received a single administration of the IL-7 protein (5 mg/kg).
  • the different treatment groups were as follows: (i) control vector only (Gl); (ii) DNA vaccine only (G2); (iii) IL-7 protein alone (G3); and (iv) DNA vaccine + IL-7 protein (G4).
  • the animals were intravenously injected with CFSE-labeled naive splenocytes that were either unpulsed or pulsed with the neoantigens. Then, 24 hours later, animals from the different treatment groups were sacrificed and percent killing of the pulsed splenocytes was assessed by measuring CFSE expression via flow cytometry.
  • EXAMPLE 6 FURTHER ANALYSIS OF ANTI-TUMOR EFFECTS OF A NUCLEOTIDE VACCINE AND IL-7 COMBINATION THERAPY [0246] Further to the anti -turn or data provided in Example 3, it was next assessed whether DNA vaccine and IL-7 combination therapy described herein can also have therapeutic effects when administered after tumor induction. Briefly, as shown in FIG. 5A, mice were implanted subcutaneously with E0771 tumor cells in each flank (see, e.g., Example 3). Once palpable tumor size was reached (between about 25-100 mm 3 ; approximately 3-6 days after tumor implantation), the animals were randomized (i.e., day 0).
  • tumor animals that were treated with the combination therapy generally had greater tumor-specific T cell immune responses compared to animals from the different treatment groups.
  • EXAMPLE 7 ANALYSIS OF A NUCLEOTIDE VACCINE AND IL-7 COMBINATION THERAPY AS A PROPHYLACTIC CANCER VACCINE
  • mice were immunized with the control vector or the DNA vaccine construct as described in Examples 1 and 2. Specifically, the mice received the DNA vaccine for a total of three doses (4 pg / dose) at a dosing frequency of once every three days (i.e., days 0, 3, and 6). See FIG. 6A. At day 13 post initial DNA vaccine administration, some of the animals received a single intravenous dose of the IL-7 protein (5 mg/kg).
  • the treatment groups were as follows: (i) control vector alone ("vector”); (ii) DNA vaccine alone ("nAg”); (iii) IL-7 protein alone ("IL-7 only”); and (iv) DNA vaccine and IL-7 ("nAg + IL-7").
  • vector control vector alone
  • nAg DNA vaccine alone
  • IL-7 protein alone
  • IL-7 + IL-7 DNA vaccine and IL-7

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EP21830359.2A 2020-11-05 2021-11-05 Verfahren zur behandlung eines tumors mit einer kombination aus einem il-7-protein und einem nukleotidimpfstoff Pending EP4240408A1 (de)

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Family Cites Families (60)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4044126A (en) 1972-04-20 1977-08-23 Allen & Hanburys Limited Steroidal aerosol compositions and process for the preparation thereof
GB1429184A (en) 1972-04-20 1976-03-24 Allen & Hanburys Ltd Physically anti-inflammatory steroids for use in aerosols
US5354844A (en) 1989-03-16 1994-10-11 Boehringer Ingelheim International Gmbh Protein-polycation conjugates
JP3485184B2 (ja) 1989-07-14 2004-01-13 アメリカン サイアナミド カンパニー インターロイキン含有安定ワクチン組成物
US5585112A (en) 1989-12-22 1996-12-17 Imarx Pharmaceutical Corp. Method of preparing gas and gaseous precursor-filled microspheres
IT1246382B (it) 1990-04-17 1994-11-18 Eurand Int Metodo per la cessione mirata e controllata di farmaci nell'intestino e particolarmente nel colon
US5543390A (en) 1990-11-01 1996-08-06 State Of Oregon, Acting By And Through The Oregon State Board Of Higher Education, Acting For And On Behalf Of The Oregon Health Sciences University Covalent microparticle-drug conjugates for biological targeting
US5643578A (en) 1992-03-23 1997-07-01 University Of Massachusetts Medical Center Immunization by inoculation of DNA transcription unit
US6010715A (en) 1992-04-01 2000-01-04 Bertek, Inc. Transdermal patch incorporating a polymer film incorporated with an active agent
US6024975A (en) 1992-04-08 2000-02-15 Americare International Diagnostics, Inc. Method of transdermally administering high molecular weight drugs with a polymer skin enhancer
US6274552B1 (en) 1993-03-18 2001-08-14 Cytimmune Sciences, Inc. Composition and method for delivery of biologically-active factors
US5523092A (en) 1993-04-14 1996-06-04 Emory University Device for local drug delivery and methods for using the same
US5985307A (en) 1993-04-14 1999-11-16 Emory University Device and method for non-occlusive localized drug delivery
US6004534A (en) 1993-07-23 1999-12-21 Massachusetts Institute Of Technology Targeted polymerized liposomes for improved drug delivery
US5759542A (en) 1994-08-05 1998-06-02 New England Deaconess Hospital Corporation Compositions and methods for the delivery of drugs by platelets for the treatment of cardiovascular and other diseases
US5660854A (en) 1994-11-28 1997-08-26 Haynes; Duncan H Drug releasing surgical implant or dressing material
US5983134A (en) 1995-04-23 1999-11-09 Electromagnetic Bracing Systems Inc. Electrophoretic cuff apparatus drug delivery system
US6316652B1 (en) 1995-06-06 2001-11-13 Kosta Steliou Drug mitochondrial targeting agents
US6167301A (en) 1995-08-29 2000-12-26 Flower; Ronald J. Iontophoretic drug delivery device having high-efficiency DC-to-DC energy conversion circuit
US6013516A (en) 1995-10-06 2000-01-11 The Salk Institute For Biological Studies Vector and method of use for nucleic acid delivery to non-dividing cells
US6039975A (en) 1995-10-17 2000-03-21 Hoffman-La Roche Inc. Colon targeted delivery system
WO1998008947A1 (en) * 1995-12-19 1998-03-05 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Enhancement of dna immunization through the use of cytokines
TW345603B (en) 1996-05-29 1998-11-21 Gmundner Fertigteile Gmbh A noise control device for tracks
US7118754B1 (en) 1996-07-30 2006-10-10 Transgene S.A. Pharmaceutical composition for treating papillomavirus tumors and infection
US5985317A (en) 1996-09-06 1999-11-16 Theratech, Inc. Pressure sensitive adhesive matrix patches for transdermal delivery of salts of pharmaceutical agents
US6139865A (en) 1996-10-01 2000-10-31 Eurand America, Inc. Taste-masked microcapsule compositions and methods of manufacture
PT904392E (pt) 1996-10-17 2001-06-29 Oxford Biomedica Ltd Vectores retrovirais
GB9621680D0 (en) 1996-10-17 1996-12-11 Oxford Biomedica Ltd Lentiviral vectors
GB9622500D0 (en) 1996-10-29 1997-01-08 Oxford Biomedica Ltd Therapeutic gene
US6131570A (en) 1998-06-30 2000-10-17 Aradigm Corporation Temperature controlling device for aerosol drug delivery
US5860957A (en) 1997-02-07 1999-01-19 Sarcos, Inc. Multipathway electronically-controlled drug delivery system
US6120751A (en) 1997-03-21 2000-09-19 Imarx Pharmaceutical Corp. Charged lipids and uses for the same
US6730512B2 (en) 1997-04-09 2004-05-04 Amdl, Inc. Combination immunogene therapy
US6060082A (en) 1997-04-18 2000-05-09 Massachusetts Institute Of Technology Polymerized liposomes targeted to M cells and useful for oral or mucosal drug delivery
US5948433A (en) 1997-08-21 1999-09-07 Bertek, Inc. Transdermal patch
EP0968711B9 (de) 1997-10-28 2008-05-28 Bando Chemical Industries, Ltd. Dermatologisches pflaster und verfahren zur herstellung seiner basisschicht
US5994136A (en) 1997-12-12 1999-11-30 Cell Genesys, Inc. Method and means for producing high titer, safe, recombinant lentivirus vectors
US6048736A (en) 1998-04-29 2000-04-11 Kosak; Kenneth M. Cyclodextrin polymers for carrying and releasing drugs
JP2002526079A (ja) 1998-09-21 2002-08-20 シェーリング コーポレイション ヒトインターロイキン−ｂ５０、治療的使用
US6271359B1 (en) 1999-04-14 2001-08-07 Musc Foundation For Research Development Tissue-specific and pathogen-specific toxic agents and ribozymes
US6256533B1 (en) 1999-06-09 2001-07-03 The Procter & Gamble Company Apparatus and method for using an intracutaneous microneedle array
US6261595B1 (en) 2000-02-29 2001-07-17 Zars, Inc. Transdermal drug patch with attached pocket for controlled heating device
US20020106798A1 (en) 2000-03-02 2002-08-08 Robinson Harriet L. DNA expression vectors and methods of use
CA2404479A1 (en) 2000-03-30 2001-10-11 University Of Connecticut Hybrid cytokine of il-7 and .beta.-chain of hepatocyte growth factor
AU2002242016A1 (en) 2001-02-01 2002-08-12 The Johns Hopkins University Nucleic acid derived vaccine that encodes an antigen linked to a polypeptide that promotes antigen presentation
EP1391513A1 (de) 2002-08-08 2004-02-25 Cytheris IL-7 als Wirkstoff, IL-7-haltige Zusammensetzung, Herstellungsverfahren und Verwendungen
US20050054054A1 (en) 2002-11-12 2005-03-10 Foss Francine M. Interleukin-7 molecules with altered biological properties
JP2008502317A (ja) 2003-12-30 2008-01-31 メルク　パテント　ゲゼルシャフト　ミット　ベシュレンクテル　ハフトング Ｉｌ−７融合タンパク質
ES2342964T3 (es) 2004-12-09 2010-07-20 Merck Patent Gmbh Variantes de la interleucina-7 con inmunogenicidad reducida.
EP1746161A1 (de) 2005-07-20 2007-01-24 Cytheris Glykosylierte IL-7, Verfahren zur Herstellung und Verwendung davon
GB2434578A (en) 2006-01-26 2007-08-01 Univ Basel Transgenic animals
CN101687933B (zh) 2007-05-30 2015-11-25 浦项工科大学校产学协力团 免疫球蛋白融合蛋白
GB0815216D0 (en) 2008-08-21 2008-09-24 Asterion Ltd Interleukin
JP5969458B2 (ja) 2010-04-09 2016-08-17 アルブミディクス アクティーゼルスカブ アルブミン誘導体及び変異体
KR101873201B1 (ko) 2015-06-11 2018-07-02 주식회사 제넥신 변형된 인터루킨-7 단백질 및 이의 용도
EP3331555A1 (de) 2015-08-05 2018-06-13 CureVac AG Epidermaler mrna-impfstoff
MX2018004916A (es) 2015-10-22 2019-07-04 Modernatx Inc Vacuna contra el virus de la influenza de amplio espectro.
KR102386735B1 (ko) 2015-11-06 2022-04-14 주식회사 제넥신 변형된 인터루킨-7 융합 단백질의 제형
WO2019151760A1 (ko) * 2018-02-02 2019-08-08 주식회사 에스엘백시젠 신규 다가 hpv 백신 조성물
AU2019219200A1 (en) * 2018-02-12 2020-08-06 BioNTech SE Treatment using cytokine encoding RNA

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